JP2018502910A - Pharmaceutical product and stable liquid composition of IL-17 antibody - Google Patents

Abstract

The present disclosure is directed to pharmaceutical products and stable liquid compositions of IL-17 antibodies and antigen-binding fragments thereof, such as AIN457 (secukinumab), and methods of making these pharmaceutical products and compositions. The present disclosure describes the (and Also intended for use (eg as part of a kit with instructions for use).

Description

This application claims priority to US Provisional Application No. 62/009467, filed Dec. 22, 2014, which is incorporated herein by reference in its entirety.

  The present disclosure is directed to pharmaceutical products comprising stable liquid pharmaceutical compositions of IL-17 antibodies and antigen-binding fragments thereof, such as AIN457 (secukinumab), and methods of making such pharmaceutical products and liquid pharmaceutical compositions. .

  IL-17A is a central lymphokine of a newly defined subset of Th17 cells, an inflammatory T cell that is important in several autoimmune and inflammatory processes. Neutralization of IL-17A is expected to treat the pathophysiology underlying immune mediated diseases, resulting in symptom relief. Sexukinumab (AIN457) is a high-affinity fully human monoclonal anti-human antibody that inhibits IL-17A activity, and it has various autoimmune diseases such as rheumatoid arthritis, ankylosing spondylitis, psoriatic arthritis, diabetes, It has emerged as a potential treatment for patients with asthma, chronic plaque psoriasis and multiple sclerosis. Several Phase II and III trials have shown that secukinumab is superior to placebo in achieving PASI 75 in the treatment of chronic plaque-type psoriasis (eg, both 3 × 150 mg and 3 × 75 mg of secukinumab in the CAIN457A2220 study). Were superior to placebo in achieving PASI 75 at 12 weeks (81.5% and 57.1% vs. 9.1%, respectively)). Sekukinumab is currently used in the global phase III trial for the treatment of chronic plaque-type psoriasis and has also been shown to be superior to placebo and newly superior to etanercept.

  International patent application PCT / EP2011 / 066946 provides a lysed composition of sucrose-based secukinumab, which is reconstituted with 1 mL of water just prior to use. However, PCT / EP2011 / 066946 does not provide a ready-to-use pharmaceutical product or liquid pharmaceutical composition disclosure of secukinumab having long-term stability. Indeed, the limit stability of proteins in liquid compositions often prevents long-term storage at room temperature or refrigerated conditions. Furthermore, various physical and chemical reactions (aggregation [covalent and non-covalent], deamidation, oxidation, clipping, isomerization, denaturation) can occur in solution, increasing the level of degradation products and / or Leading to loss of biological activity. To ensure that dosage and product safety claims are met when the molecule is administered to a patient, commercially available ready-to-use liquid antibody compositions provide sufficient physical and chemical properties of the antibody during shipping and handling. Should provide stability. Specifically, acceptable liquid antibody compositions must enhance stability and minimize proteolysis, particularly protein aggregation, in order to avoid significant immunogenic reactions. In addition, the composition must be of an osmolarity and pH value that is acceptable for subcutaneous application, and must also have a low viscosity as a requirement for manufacture (mixing, filtration, filling) and needle passage. I must. Since it is difficult to balance these myriad requirements, the production of commercially realistic aqueous biopharmaceutical compositions has become a technical challenge.

  In spite of the technical challenges outlined above, the inventors have identified new beneficial ready-to-use pharmaceutical products and liquid pharmaceutical compositions of the IL-17 antibodies and antigen-binding fragments disclosed herein. For example, sexualkinumab has now been successfully developed.

The present disclosure provides containers (eg, pens, syringes, vials) having a headspace with less than about 12% oxygen (eg, less than about 10% oxygen, less than about 8% oxygen, less than about 6% oxygen, etc.). , An autoinjector) and a liquid product disposed in a container. The liquid composition is a ready-to-use liquid composition that is not reconstituted from a lyophilizate and is generally at least one of the disclosed IL-17 antibodies or antigen-binding fragments thereof (eg, secukinumab), a buffering agent , Surfactants, methionine and stabilizers, and sub-combinations thereof. The combined use of certain stabilizers and low oxygen levels in the container headspace significantly contributes to the long-term stability of the liquid pharmaceutical product and reduces the oxidation of IL-17 antibodies (eg, secukinumab) contained in the composition. The inventors determined that it would be blocked. These liquid compositions have excellent properties, for example:
Aggregates of no more than 3.5% with 2.5 mM methionine, no more than 3.0% with 5 mM; and no more than 2.2% with a composition containing 20 mM methionine, as measured by SEC after 13 months storage at 25 ° C. Formation; and after storage at 25 ° C. for 13 months, by RP-HPLC (sum of the mutants before the main peak) 2.5 mM at 39.4% or less, 5.0 mM at 37.8% or less and Decomposition product of 34.5% or less with a composition containing 20 mM methionine.

  Accordingly, herein, a container having a headspace in which the oxygen content is less than about 12%, as well as a liquid medicament having a pH of about 5.2 to about 6.2 disposed in said container A pharmaceutical product comprising a liquid pharmaceutical composition comprising about 20 mg / ml to about 175 mg / ml secukinumab and about 2.5 to about 20 mM L-methionine, which has not been reconstituted from a lyophilizate. Disclosed.

  As used herein, a container having a headspace in which the oxygen content is less than about 6%, as well as a liquid pharmaceutical composition disposed in said container, comprising about 25 mg / mL to about 150 mg / mL IL-17 antibody disclosed herein (eg, secukinumab), about 10 mM to about 30 mM histidine pH 5.8, about 200 mM to about 225 mM trehalose, about 0.02% polysorbate 80 and about 2.5 mM to about 20 mM methionine And a pharmaceutical product comprising a liquid pharmaceutical composition that has not been reconstituted from a lyophilizate.

  As used herein, an IL-17 antibody (eg, secukinumab) disclosed herein having a pH of about 5.2 to about 6.2 and having a pH of about 25 mg / ml to about 150 mg / ml, and about 2 Preparing a liquid composition comprising 5 mM to about 20 mM methionine; placing the liquid composition in a container having a headspace; and adjusting the oxygen content in the headspace to about 12% or less. Also disclosed is a method for reducing the oxidation of secukinumab.

  As used herein, from about 25 mg / mL to about 150 mg / mL of an IL-17 antibody disclosed herein (eg, secukinumab), from about 10 mM to about 30 mM buffer (eg, histidine) pH 5.8, about Also disclosed is a stable liquid pharmaceutical composition comprising 200 mM to about 225 mM stabilizer (eg, trehalose), about 0.02% surfactant (eg, polysorbate 80) and about 2.5 mM to about 20 mM methionine. The

  The present disclosure provides these pharmaceutical products and stable liquid compositions for the treatment of various IL-17 mediated disorders such as autoimmune disorders such as psoriasis, ankylosing spondylitis, psoriatic arthritis and rheumatoid arthritis. As well as kits containing these pharmaceutical products and stable liquid compositions.

  Additional compositions, products, methods, regimens, uses and kits are provided in the following description and the appended claims. Further features, advantages and aspects of the present disclosure will become apparent to those skilled in the art from the following description and appended claims.

FIGS. 1A-D show the effect of different antioxidant stabilizers on the stability of 150 mg / ml secukinumab syringe solution: 1 μm subvisible particles by light darkening after 8 weeks at 5 ° C. (A) Pre-main peak species (%) by RP-HPLC after 8 weeks at 25 ° C. (B) DP-SEC (%) after 8 weeks at 40 ° C. (C) ) AP-SEC (%) after 8 weeks at 40 ° C. (D) FIG. 5 shows the effect of L-methionine concentration on the stability of 25 mg / ml secukinumab when stored at 25 ° C .: Premain peak species (%) by RP-HPLC Gray dashed line: 10 mM L-methionine / 5 Linear fit to% headspace oxygen content data; black dashed line: linear fit to 0 mM L-methionine / 5% headspace oxygen content data; FIG. 5 shows the effect of L-methionine, trehalose and polysorbate 80 on the stability of 150 mg / ml secukinumab syringe solution stored at 25 ° C. for 6 months: pre-main peak species (%) by RP-HPLC. (FIG. 4A and B) It is a figure which shows the influence of the L-methionine density | concentration on the stability of the 150 mg / ml secukinumab syringe solution preserve | saved at 5 degreeC. AP-SEC (%) in the presence of 5 mM and 0 mM L-methionine (A) and pre-main peak species (%) by RP-HPLC (B). It is a figure which shows the influence of L-methionine density | concentration on the stability of a 150 mg / ml secukinumab syringe liquid solution after 30 months at 5 degreeC and 13 months at 25 degreeC. Pre-main peak species (%) by AP-SEC (%) (A) and RP-HPLC (B). FIGS. 6A and B show the effect of L-methionine concentration on the stability of 25 mg / ml secukinumab vials (10% headspace oxygen content) after storage at 40 ° C. for 3 months. Total impurities (%) by AP-SEC (%) (A) and CE-SDS (non-reduced) (B). FIG. 2 shows the effect of headspace oxygen content on 150 mg / ml secukinumab syringe solution stored at 25 ° C .: AP-SEC (%). (FIGS. 8A-D) Headspace oxygen content and loading on AP-SEC (%) in 150 mg / ml secukinumab syringe solution after storage at 25 ° C. (A, B) and 5 ° C. (C, D). It is a figure which shows the influence of a volume. The filling volume of A and C is 0.5 mL. The filling volume of B and D is 1.0 mL. FIG. 5 shows the effect of headspace oxygen content and fill volume on the stability of 150 mg / ml secukinumab syringe solution after 6 months at 5 ° C. and 25 ° C .: purity by RP-HPLC. FIG. 2 shows the effect of L-methionine concentration and headspace oxygen content on the stability of 150 mg / ml secukinumab syringe solution stored at 25 ° C. for 6 months (A): AP-SEC (%); FIG. 5 shows the effect of nitrogen purge and L-methionine concentration on the stability of 150 mg / ml secukinumab syringe solution: pre-main peak species (%) by RP-HPLC. (FIGS. 12A and B) FIG. 12 shows the effect of pH on the stability of 150 mg / ml secukinumab syringe solution after storage at 40 ° C. for 4 weeks. Scaled estimate for stability / 2 (effect of increasing pH by 0.3 units). Pre-main peak species by RP-HPLC (% change) (A) and AP-SEC (% change) (B). FIGS. 13A-D show the effect of pH on the stability of 150 mg / ml secukinumab syringe solution after storage at 5 ° C .: Turbidity (NTU) (A), purity by SEC (%) (B) Acidic variant by CEX (%) (C), AP-SEC (%) (D). FIG. 2 shows the effect of stabilizers on the stability of 150 mg / ml secukinumab syringe solution after 8 weeks storage at 25 ° C .: AP-SEC parameter estimates. FIG. 2 shows the effect of surfactant on the stability of 150 mg / ml secukinumab syringe solution after shaking: parameter estimates (subparticle number / ml) for sub-visible particles ≧ 1 μm due to light darkening. FIGS. 16A-D show the effect of buffer type on the stability of 150 mg / ml secukinumab syringe solution: AP-SEC parameter estimates (%) after freeze-thaw stress (A) ), AP-SEC (%) after shaking stress (B), Pre-main peak species (%) by RP-HPLC after storage at 25 ° C. for 8 weeks (C), DP-SEC (%) after shaking stress ) (D).

  The term “comprising” encompasses “comprising” and “consisting of”, for example, a composition “comprising” X can consist exclusively of X or can include additional ones, for example X + Y.

  Unless the context dictates otherwise, the term “about” with respect to a numerical value x means +/− 10%.

  “Monthly” means about every 4 weeks (eg, every 4 weeks), which is about every 28 days (eg, every 28 days).

The term “antibody” as referred to herein includes intact antibodies and any antigen binding fragment or single chain thereof. Naturally occurring “antibodies” are glycoproteins comprising at least two heavy (H) chains and two light (L) chains interconnected by disulfide bonds. Each heavy chain is comprised of a heavy chain variable region (abbreviated herein as V _H ) and a heavy chain constant region. The heavy chain constant region is comprised of three domains, CH1, CH2 and CH3. Each light chain is comprised of a light chain variable region (abbreviated herein as VL) and a light chain constant region. The light chain constant region is comprised of one domain, CL. The _VH and _VL regions are further subdivided into hypervariable regions called hypervariable regions or complementarity determining regions (CDRs) interspersed with more conserved regions called framework regions (FR) can do. Each _VH and _VL is composed of three CDRs and four FRs arranged in the following order from the amino terminus to the carboxy terminus: FR1, CDR1, FR2, CDR2, FR3, CDR3, FR4. The variable region of the heavy and light chains contains a binding domain that interacts with an antigen. The constant region of the antibody mediates immunoglobulin binding to host tissues or factors, including various cells of the immune system (eg, effector cells) and the first component of the classical complement system (C1q). Can do. In some embodiments of the disclosed methods, regimens, kits, processes, uses and compositions, an antibody to IL-17 or IL-17 receptor, preferably an antibody to IL-17, such as secukinumab, is used. .

  As used herein, an “isolated antibody” refers to an antibody that is substantially free of other antibodies with different antigen specificities (eg, an isolated antibody that specifically binds IL-17. The antibody has substantially no antibody that specifically binds to an antigen other than IL-17). The terms “monoclonal antibody” or “monoclonal antibody composition” as used herein refer to a preparation of antibody molecules of single molecular composition. As used herein, the term “human antibody” is intended to include antibodies having variable regions in which both the framework and CDR regions are derived from sequences of human origin. “Human antibodies” need not be produced by humans, human tissues or human cells. The human antibodies of the present disclosure can include amino acid residues that are not encoded by human sequences (eg, in vitro random or site-directed mutagenesis, such as by in vitro conjugation during antibody gene recombination). Mutations introduced by N-nucleotide addition or by somatic mutation in vivo). In some embodiments of the disclosed methods, regimens, kits, processes, uses and compositions, the IL-17 antibody is a human antibody, an isolated antibody and / or a monoclonal antibody.

As used herein, the term “antigen-binding fragment” of an antibody refers to a fragment of an antibody that retains the ability to specifically bind to an antigen (eg, IL-17). It has been found that the antigen-binding function of an antibody can be performed by fragments of a full-length antibody. Examples of binding fragments encompassed by the term “antigen-binding fragment” of an antibody include Fab fragments that are monovalent fragments consisting of V _L , V _H , CL and CH1 domains; linked by disulfide bridges at the hinge region. F (ab) 2 fragment, which is a bivalent fragment comprising two Fab fragments; an Fd fragment consisting of the V _H and CH1 domains; an Fv fragment consisting of the V _L and V _H domains of a single arm of the antibody; V _H A dAb fragment consisting of a domain (Ward et al., 1989 Nature 341: 544-546); as well as an isolated CDR are included. Exemplary antigen binding sites include the secukinumab CDRs shown in SEQ ID NOs: 1-6 and 11-13 (Table 1), preferably the heavy chain CDR3. In addition, the two domains of the Fv fragment, _VL and _VH, are encoded by separate genes, which use recombinant methods to form a monovalent molecule in which the _VL and _VH regions are paired. A single protein chain (known as single chain Fv (scFv); see, for example, Bird et al., 1988 Science 242: 423-426; and Huston et al., 1988 Proc. Natl. Acad. Sci. 85 : 5879-5883) can be linked by a synthetic linker that allows them to be made. Such single chain antibodies are also intended to be encompassed within the term “antibody”. Single chain antibodies and antigen-binding fragments are obtained using conventional techniques known to those skilled in the art. In some embodiments of the disclosed methods, regimens, kits, processes, uses and compositions, single-chain antibodies or antigen-binding fragments of antibodies to IL-17 (eg, secukinumab) or IL-17 receptor are used. It is done.

  The term “pharmaceutical product” means a container (eg, pen, syringe, bag, pump, etc.) having a pharmaceutical composition disposed in the container. “Container” means any means for holding a liquid pharmaceutical composition, such as pens, syringes, vials, auto-injectors, patches, and the like. Each container has a “headspace”, ie, an area within the container that does not contain a liquid pharmaceutical composition. This headspace contains a mixture of gases such as oxygen and other gases normally found in air. The level of oxygen in the headspace can be adjusted, for example, by introducing an inert gas (eg, nitrogen, argon, etc.) into the headspace instead of oxygen. This can be accomplished either actively, for example by purging, or passively, for example by placing a container in the system and removing oxygen (eg, by vacuum, etc.). For example, purging with an inert gas, preferably nitrogen, can occur prior to, during, or during the filling of the composition into the container. As used herein, the term “oxygen content in the headspace” refers to the percentage of oxygen found in the headspace of a given container.

  A “stable” composition is one in which the protein therein substantially retains its stability (eg, physical, chemical and / or biological activity) after storage. Various analytical techniques for measuring protein stability are available in the art, including Peptide and Protein Drug Delivery, 247-301, Vincent Lee Ed., Marcel Dekker, Inc., New York, NY, Pubs. (1991 ) And Jones, A. Adv. Drug Delivery Rev. 10: 29-90 (1993). Stability can be measured for a selected time at a selected temperature. A “stable” liquid antibody composition has a refrigeration temperature (2-8 ° C.) of at least 6 months, 12 months, preferably 2 years and more preferably 3 years; or at room temperature (23-27 ° C.) No significant change is observed for 3 months, preferably 6 months and more preferably 1 year; or at least 1 month, preferably 3 months and more preferably 6 months under stress conditions (about 40 ° C.) Liquid antibody composition. Various stability criteria, eg, 10% or less, preferably 5% or less of the antibody monomer is degraded (eg as measured by SEC purity, RP-HPLC purity, CEX purity, CE-SDS purity (non-reducing), etc.) )), Can be used. Alternatively, stability can be shown if the solution remains clear to slightly opalescent by using visual analysis or nephelometry. Alternatively, stability is exhibited when the concentration, pH and osmolarity of the composition have a variation of not more than +/− 10% over a given period of time, for example at least 3 months, preferably 6 months and more preferably 1 year. be able to. Alternatively, potency (eg, as measured by biological activity, such as inhibition or CEX assay) is 70% of the control over a given period of time, eg, at least 3 months, preferably 6 months, more preferably 1 year. In the range of ~ 130% (eg, at least 70%, at least 75%, at least 76%, at least 80%, at least 90%, at least 91%, at least 95%), preferably in the range of 80-120% , Can show stability. Alternatively, clipping of 10% or less, preferably 5% or less of the antibody is observed over a given period of time, for example at least 3 months, preferably 6 months, more preferably 1 year (e.g. measured by DP-SEC etc. If you can) show stability. Alternatively, less than 10%, preferably less than 5% aggregates are formed over a given period of time, for example at least 3 months, preferably 6 months, more preferably 1 year (e.g. measured by AP-SEC etc. Sometimes) can show stability. Alternatively, if the aggregate formation as measured by SEC is ≦ about 3.5%, ≦ about 3.0%, or ≦ about 2.2% after 13 months storage at 25 ° C. Can show. Alternatively, after 13 months storage at 25 ° C., degradation product formation (as measured by RP-HPLC (pre-main peak species)) is ≦ about 39.4%, ≦ about 37.8%, or If ≦ about 34.5%, stability can be shown.

  Of aggregation, precipitation and / or denaturation as measured by visual inspection of color and / or transparency (turbidity) or by UV light scattering, size exclusion chromatography (SEC) and dynamic light scattering (DLS) If it does not show a significant increase, the antibody retains its physical stability in the pharmaceutical composition. Furthermore, the protein conformation should not be significantly altered as assessed by, for example, fluorescence spectroscopy (determining protein tertiary structure) or FTIR spectroscopy (determining protein secondary structure).

  If it does not show significant chemical modification, the antibody retains its chemical stability in the pharmaceutical composition. Chemical stability can be assessed by detecting and quantifying chemically modified forms of the protein. Degradation processes that often alter protein chemical structure include hydrolysis or clipping (assessed by methods such as size exclusion chromatography [SEC] and SDS-PAGE), oxidation (eg, combined with mass spectrometry or MALDI / TOF / MS) Evaluated by methods such as peptide mapping), deamidation (evaluated by methods such as cation exchange chromatography (CEX), capillary isoelectric focusing, peptide mapping, isoaspartic acid measurement) and isomerization ( It is evaluated by measurement of isoaspartic acid content, peptide mapping, etc.).

  An antibody retains its biological activity in a pharmaceutical composition if the biological activity of the protein / antibody for a given time is within the predetermined range of biological activity indicated at the time of preparation of the pharmaceutical composition To do. The biological activity of the antibody can be determined, for example, by the ability of an IL-17 antibody (eg, secukinumab) to bind to an antigen binding ELISA assay, potency assay (eg, bind to IL-17 and inhibit IL-6 release from chondrocytes). Or cysteamine-CEX derivatization.

  As used herein, “purity by RP-HPLC” refers to the percentage of the main peak on RP-HPLC and can be used to assess the stability of secukinumab. RP-HPLC is used to separate secukinumab and its variants by their hydrophobicity. The pre-main peak species by RP-HPLC is the percentage sum of the peaks eluting before the main peak and can contain antibody fragmented, isomerized and oxidized species.

  As used herein, “purity by CEX” refers to the percentage of the main peak in CEX and can be used to assess the stability of secukinumab antibodies. CEX is used to assess the charge heterogeneity of secukinumab by measuring the percentage of acidic and basic variants.

  As used herein, “purity by SEC” refers to the percentage of monomer at SEC and can be used to assess the stability of secukinumab. SEC is used to separate monomeric secukinumab from aggregates and fragments by their size under non-denaturing conditions. The sum of the peaks eluting before the main peak is reported as a percentage of aggregated product (AP-SEC) and the sum of the peaks eluting after the main peak is reported as a percentage of degradation product (DP-SEC).

  As used herein, “purity by CE-SDS” refers to the percentage of intact antibody in CE-SDS and can be used to assess the stability of secukinumab. CE-SDS is used to separate by-products and degradation products from intact secukinumab by their molecular size under non-reducing conditions. The sum of the peaks separated from the main peak is reported as a percentage of impurities.

  As used herein, the phrase “liquid pharmaceutical composition” is not reconstituted from a lyophilizate and includes at least one IL-17 antibody or antigen-binding fragment thereof (eg, secukinumab) and at least one additional excipient (eg, , A buffering agent). Liquid pharmaceutical compositions can contain additional excipients (stabilizers, surfactants) and additional active ingredients. This type of formulation is also referred to as a “ready to use” formulation.

  As used herein, the term “lyophilizate” refers to a dried (eg, freeze-dried) pharmaceutical composition that is substantially free of water. Antibody lyophilization techniques are well known in the art, see, eg, Rey & May (2004) Freeze-Drying / Lyophilization of Pharmaceutical & Biological Products ISBN 0824748689. Since reconstituted lyophilizate tends to have a limited shelf life, the lyophilizate is usually reconstituted to give an aqueous composition for immediate use (eg, within 1-10 days).

  The term “high concentration” refers to a composition containing more than 50 mg / ml of an antibody or antigen-binding fragment thereof. In preferred embodiments, the high concentration liquid composition is ≧ about 50 mg / ml, ≧ about 75 mg / ml, ≧ about 100 mg / ml, ≧ about 125 mg / ml, ≧ about 150 mg / ml, ≧ about 175 mg / ml, ≧ about Contains 200 mg / ml or ≧ about 225 mg / ml.

  The term “IL-17” refers to IL-17A, formerly known as CTLA8, and wild type IL-17A, IL-17A polymorphic mutations from various species (eg, human, mouse and monkey) Body and functional equivalents of IL-17A. A functional equivalent of IL-17A according to the present disclosure is preferably at least about 65%, 75%, 85%, 95%, 96%, 97%, 98 and wild type IL-17A (eg, human IL-17A). % Or even 99% overall sequence identity and substantially retain the ability to induce IL-6 production by human dermal fibroblasts.

The term “K _D ” is intended to refer to the dissociation rate of a particular antibody-antigen interaction. As used herein, the term “K _D ” shall refer to the dissociation constant obtained from the ratio of K _d to K _a (ie, K _d / K _a ) and expressed in molarity (M). . K _D values for antibodies can be determined using methods established in the art. Method for determining the K _D of an antibody is by using surface plasmon resonance, or Biacore using a biosensor system such as (R) system. In some embodiments, IL-17 antibody or antigen-binding fragment thereof binds to human IL-17 with a _{K D} of about 100~250pM (Biacore (as measured by TM)).

  The term “affinity” refers to the strength of interaction between an antibody and an antigen at a single antigenic site. Within each antigenic site, the variable region of the antibody “arm” interacts with the antigen at multiple sites via weak non-covalent forces, the more interaction the stronger the affinity. Standard assays for assessing antibody binding affinity to various species of IL-17 are known in the art, including, for example, ELISA, Western blot and RIA. The binding kinetics (eg, binding affinity) of an antibody can also be tested by standard assays known in the art, for example, Biacore® analysis.

  As used herein, the terms “subject” and “patient” include any human or non-human animal. The term “non-human animals” includes all vertebrates, eg, mammals and non-mammals, eg, non-human primates, sheep, dogs, cats, horses, cows, chickens, amphibians, reptiles, and the like.

  These IL-17 functional properties known in the art and determined by the methods described herein (eg, biochemical, immunochemical, cellular, physiological or other biological activity) An antibody that “inhibits” one or more of a particular activity statistically compared to that seen in the absence of the antibody (or in the presence of a control antibody of irrelevant specificity) It is understood that it relates to a significant decrease. Antibodies that inhibit IL-17 activity affect, for example, a statistically significant decrease of at least about 10%, at least 50%, 80%, or 90% of the parameter being measured, and the disclosed methods, uses, processes, In certain embodiments of the kits and compositions, the IL-17 antibody used can inhibit more than 95%, 98% or 99% of the functional activity of IL-17.

  As used herein, “inhibits IL-6” refers to the ability of an IL-17 antibody (eg, secukinumab) to reduce IL-6 production from chondrocytes. Biological activity of IL-17 antibodies or antigen-binding fragments thereof, such as secukinumab, inhibits IL-17-induced release of IL-6 from immortalized human chondrocyte cell lines, such as C-20 / A4 Can be measured based on its ability to do. Briefly, on the first day of the assay, C-20 / A4 cells are seeded and allowed to attach to 96-well plates and then fixed at a submaximal concentration of IL-17 (eg, about 20-200 ng in culture medium). / ML, eg about 80 ng / mL) and various concentrations of antibodies (eg about 0.01 μg / mL to about 4 μg / mL, eg about 0.5 μg / mL to about 2 μg / mL in the assay plate) Incubate overnight. To increase the dynamic range of the assay, TNFα that promotes IL-17 production induced by IL-17 is included (eg, from about 0.01 ng / mL to about 1 ng / mL, eg, about 0.1. 5 ng / mL). On day 2, the concentration of IL-6 in the cell supernatant is quantified by ELISA. The amount of IL-6 in the cell supernatant is inversely proportional to the activity of the IL-17 antibody present in the sample. The biological activity of the antibody test sample is quantified by comparing its ability to inhibit IL-17 dependent release of IL-6 to that of an antibody reference standard. Samples and standards are normalized based on protein content. Relative potency is calculated using a parallel line assay by the European Pharmacopoeia. Final results are expressed as the relative potency (percentage) of the sample compared to the reference standard.

Unless otherwise indicated, the term “derivative” refers to, for example, an amino acid sequence variant and a covalent modification of an IL-17 antibody or antigen-binding fragment thereof according to the present disclosure, such as secukinumab, of a specified sequence (eg, a variable domain). For example, pegylation, deamidation, hydroxylation, phosphorylation, methylation, etc.). “Functional derivatives” include molecules having qualitative biological activity in common with the disclosed IL-17 antibodies or antigen-binding fragments thereof. Functional derivatives include fragments and peptide analogs of the IL-17 antibody or antigen-binding fragments thereof disclosed herein. A fragment includes a region within the sequence of a polypeptide according to the present disclosure, eg, of a specified sequence. A functional derivative of an IL-17 antibody or antigen-binding fragment thereof disclosed herein (eg, a functional derivative of secukinumab) is a V _H and / or V of the IL-17 binding molecule disclosed herein. _At least about 65%, 75%, 85%, 95%, 96%, 97%, 98%, or even 99% overall sequence identity with the _L sequence (eg, the _VH and / or _VL sequences of Table 1) V _H and / or V that have sex and substantially retain the ability to bind to human IL-17, or inhibit IL-6 production of human skin fibroblasts induced by, for example, IL-17 _{The L} domain is preferably included.

The phrase “substantially identical” means that the related amino acid or nucleotide sequences (eg, V _H or V _L domains) are identical or have substantive differences compared to a particular reference sequence ( For example, through conserved amino acid substitutions). Non-substantial differences include minor amino acid changes, such as one or two substitutions in the five amino acid sequences of a specified region (eg, V _H or _VL domain). In the case of an antibody, the second antibody has the same specificity and at least 50% of its affinity. Sequences that are substantially identical to the sequences disclosed herein (eg, at least about 85% sequence identity) are also part of this application. In some embodiments, the sequence identity of the derivative IL-17 antibody (eg, a derivative of secukinumab, eg, secukinumab biologically similar antibody) is about 90% or greater, eg, 90%, to the disclosed sequence, It may be 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more.

  “Identity” with respect to a native polypeptide and functional derivatives thereof, as used herein, refers to any conservative after aligning sequences to achieve the maximum percent identity, and introducing gaps as necessary. Substitutions are also defined as the percentage of amino acid residues in the candidate sequence that are identical to the corresponding natural polypeptide residue when not considered part of sequence identity. None of N-terminal or C-terminal extensions and insertions shall be construed as reducing identity. Methods and computer programs for alignment are well known. Percent identity is calculated using standard alignment algorithms such as the Basic Local Alignment Search Tool (BLAST) described by Altshul et al. ((1990) J. Mol. Biol., 215: 403 410); Needleman et al. ((1970) J Mol. Biol., 48: 444 453); or by the algorithm of Meyers et al. ((1988) Comput. Appl. Biosci., 4:11 17). The parameter set may be a Blosum62 scoring matrix with 12 gap penalties, 4 gap extension penalties, and 5 frameshift gap penalties. The percent identity between two amino acid or nucleotide sequences is determined by the E.I. Meyers and W.M. The algorithm of Miller ((1989) CABIOS, 4: 11-17) can also be used to determine using the PAM120 weight residue table, a gap length penalty of 12, and a gap penalty of 4.

  “Amino acid (s)” refers to, for example, all naturally occurring L-α-amino acids and includes D amino acids. The phrase “amino acid sequence variant” refers to molecules that have some difference in their amino acid sequences when compared to sequences according to the present disclosure. A polypeptide according to the present disclosure, for example an amino acid sequence variant of a specified sequence, binds to human IL-17 or inhibits IL-6 production in human dermal fibroblasts induced by, for example, IL-17 Still have. Amino acid sequence variants are substitutional variants (where at least one amino acid residue is removed and instead a different amino acid is inserted at the same position in a polypeptide according to the present disclosure), insertion variants (according to the present disclosure). A polypeptide in which one or more amino acids are inserted immediately next to the amino acid at a particular position in the polypeptide), and a deletion variant (one in which one or more amino acids are removed in a polypeptide according to the present disclosure). Including.

  The term “pharmaceutically acceptable” means a non-toxic material that does not interfere with the effectiveness of the biological activity of the active ingredient (s).

  The term “administering” with respect to a compound, eg, an IL-17 binding molecule or another agent, is used to refer to delivering the compound to the patient by any route.

  As used herein, a “therapeutically effective amount” is the treatment, prevention, prevention of its onset of at least one symptom of a disorder or recurrent disorder by administration of a single or multiple doses to a patient (eg, a human). IL-17 antibody or antigen-binding fragment thereof that is effective in healing, delaying, reducing its severity, ameliorating, or extending patient survival beyond what would be expected in the absence of such treatment For example, refers to the amount of secukinumab. When applied to an individual active ingredient administered alone (eg, an IL-17 antibody, eg, secukinumab), the term refers only to that ingredient. When applied to a combination, the term refers to the combined amount of active ingredients that provide a therapeutic effect, whether administered together, sequentially or simultaneously.

  The term “treatment” or “treating” refers to both prophylactic or preventative treatment as well as therapeutic or disease modifying treatment, in patients at risk of becoming or suspected of becoming ill, and in illness. Includes treatment of patients diagnosed as having or suffering from a disease or medical condition, as well as suppression of clinical recurrence. Treatment exceeds what would be expected to prevent, cure, delay its onset, reduce or improve severity of, or in the absence of such treatment, one or more symptoms of a disorder or recurrent disorder It can be given to patients who have a medical disorder or who may eventually get the disorder to prolong the survival of the patient.

  The phrase “means for administering” refers to any available device for systemically administering a drug to a patient, such as, but not limited to, pre-filled syringes, vials and syringes, infusion pens, automatic infusion devices, i . v. Used to indicate infusions and bags, pumps, patch pumps, etc. Such items allow the patient to self-administer the drug (ie, administer the drug on their own) or allow the physician to administer the drug. In general, the dosage given in “mg / kg” is i. v. The dose administered through the route and given in “mg” is i. m. Or s. c. Administered through injection. In some embodiments of the disclosed methods, kits, regimens and uses, the IL-17 antibody or antigen-binding fragment thereof, eg, secukinumab, is i. v. Delivered to the patient via the route. In some embodiments of the disclosed methods, kits, regimens and uses, the IL-17 antibody or antigen-binding fragment thereof, eg, secukinumab, is s. c. Delivered to the patient via the route.

IL-17 Antibodies and Antigen Binding Fragments thereof Disclosed pharmaceutical products, compositions, liquid compositions, regimens, processes, uses, methods and kits contain or utilize IL-17 antibodies or antigen binding fragments thereof.

In one embodiment, the IL-17 antibody or antigen-binding fragment thereof, eg, secukinumab, comprises at least one immunoglobulin heavy chain variable domain (V _H ) comprising hypervariable region CDR1, CDR2, and CDR3, wherein CDR1 is SEQ ID NO: The CDR2 has the amino acid sequence of SEQ ID NO: 2, and the CDR3 has the amino acid sequence of SEQ ID NO: 3. In one embodiment, the IL-17 antibody or antigen-binding fragment thereof, eg secukinumab, comprises at least one immunoglobulin light chain variable domain (V _L ) comprising hypervariable regions CDR1 ′, CDR2 ′ and CDR3 ′, wherein said CDR1 'Has the amino acid sequence of SEQ ID NO: 4, the CDR2' has the amino acid sequence of SEQ ID NO: 5, and the CDR3 'has the amino acid sequence of SEQ ID NO: 6. In one embodiment, the IL-17 antibody or antigen-binding fragment thereof, eg secukinumab, comprises at least one immunoglobulin heavy chain variable domain (V _H ) comprising hypervariable regions CDR1-x, CDR2-x and CDR3-x. The CDR1-x has the amino acid sequence of SEQ ID NO: 11, the CDR2-x has the amino acid sequence of SEQ ID NO: 12, and the CDR3-x has the amino acid sequence of SEQ ID NO: 13.

In one embodiment, the IL-17 antibody or antigen-binding fragment thereof, eg, secukinumab, comprises at least one immunoglobulin V _H domain and at least one immunoglobulin _VL domain, and a) the immunoglobulin V _H domain is (eg, I) CDR1 has the amino acid sequence of SEQ ID NO: 1, CDR2 has the amino acid sequence of SEQ ID NO: 2, CDR3 has the amino acid sequence of SEQ ID NO: 3, hypervariable regions CDR1, CDR2 and CDR3 Or ii) CDR1-x has the amino acid sequence of SEQ ID NO: 11, CDR2-x has the amino acid sequence of SEQ ID NO: 12, and CDR3-x has the amino acid sequence of SEQ ID NO: 13, x, CDR2-x and CDR3-x, b) an immunoglobulin _VL domain (e.g. For example, hypervariable regions CDR1 ′, CDR2 ′ and CDR3 ′, wherein CDR1 ′ has the amino acid sequence of SEQ ID NO: 4, CDR2 ′ has the amino acid sequence of SEQ ID NO: 5, and CDR3 ′ Has the amino acid sequence of SEQ ID NO: 6.

In one embodiment, the IL-17 antibody or antigen-binding fragment thereof, eg secukinumab, is a) an immunoglobulin heavy chain variable domain (V _H ) comprising the amino acid sequence set forth in SEQ ID NO: 8; b) the amino acid sequence set forth in SEQ ID NO: 10. An immunoglobulin light chain variable domain (V _L ) comprising: c) an immunoglobulin V _H domain comprising the amino acid sequence set forth in SEQ ID NO: 8 and an immunoglobulin _VL domain comprising the amino acid sequence set forth in SEQ ID NO: 10; d) SEQ ID NO: 1 E) an immunoglobulin V _H domain comprising the hypervariable region shown in SEQ ID NO: 2 and SEQ ID NO: 3; e) an immunoglobulin _VL domain comprising the hypervariable region shown in SEQ ID NO: 4, SEQ ID NO: 5 and SEQ ID NO: 6; f) sequence No. 11, an immunoglobulin _{V H} domain comprising the hypervariable regions as shown in SEQ ID NO: 12 and SEQ ID NO: 13; g) SEQ ID 1, the immunoglobulin V _H domain and SEQ ID NO: 4, an immunoglobulin V _L domain comprising the hypervariable regions as shown in SEQ ID NO: 5 and SEQ ID NO: 6 comprising the hypervariable regions as shown in SEQ ID NO: 2 and SEQ ID NO: 3; or h) sequences An immunoglobulin V _H domain comprising the hypervariable regions shown in SEQ ID NO: 11, SEQ ID NO: 12 and SEQ ID NO: 13 and an immunoglobulin _VL domain comprising the hypervariable regions shown in SEQ ID NO: 4, SEQ ID NO: 5 and SEQ ID NO: 6.

  For ease of reference, the amino acid sequence of the hypervariable region of the secukinumab monoclonal antibody, based on the Kabat definition and determined by X-ray analysis, and using the Chothia and collaborator approach, is shown in Table 1 below. To provide.

  In a preferred embodiment, the constant region domain is a suitable human constant, as described, for example, in “Sequences of Proteins of Immunological Interest”, Kabat EA et al, US Department of Health and Human Services, Public Health Service, National Institute of Health. A region domain is also preferably included. The DNA encoding Secukinumab VL is shown in SEQ ID NO: 9. The DNA encoding secukinumab VH is shown in SEQ ID NO: 7.

  In some embodiments, the IL-17 antibody or antigen-binding fragment thereof, eg, secukinumab, comprises three CDRs of SEQ ID NO: 10. In other embodiments, the IL-17 antibody comprises three CDRs of SEQ ID NO: 8. In other embodiments, the IL-17 antibody comprises three CDRs of SEQ ID NO: 10 and three CDRs of SEQ ID NO: 8. The CDRs of SEQ ID NO: 8 and SEQ ID NO: 10 as defined by Chothia and Kabat can be found in Table 1.

  In some embodiments, the IL-17 antibody or antigen-binding fragment thereof, eg, secukinumab, comprises the light chain of SEQ ID NO: 14. In other embodiments, the IL-17 antibody or antigen-binding fragment thereof, eg, secukinumab, comprises the heavy chain of SEQ ID NO: 15. In other embodiments, the IL-17 antibody or antigen-binding fragment thereof, eg, secukinumab, comprises a light chain of SEQ ID NO: 14 and a heavy chain of SEQ ID NO: 15. In some embodiments, the IL-17 antibody or antigen-binding fragment thereof, eg, secukinumab, comprises three CDRs of SEQ ID NO: 14. In other embodiments, the IL-17 antibody or antigen-binding fragment thereof, eg, secukinumab, comprises three CDRs of SEQ ID NO: 15. In other embodiments, the IL-17 antibody or antigen-binding fragment thereof, eg, secukinumab, comprises 3 CDRs of SEQ ID NO: 14 and 3 CDRs of SEQ ID NO: 15. The CDRs of SEQ ID NO: 15 and SEQ ID NO: 17 as defined by Chothia and Kabat can be found in Table 1.

  The hypervariable region may bind to any type of framework region, but is preferably of human origin. Suitable framework regions are described in Kabat E.A. et al, ibid. A preferred heavy chain framework is a human heavy chain framework, such as that of a secukinumab antibody. For example, FR1 (amino acids 1 to 30 of SEQ ID NO: 8), FR2 (amino acids 36 to 49 of SEQ ID NO: 8), FR3 (amino acids 67 to 98 of SEQ ID NO: 8) and FR4 (amino acids 117 to 8 of SEQ ID NO: 8) in this order. 127) area. Considering the hypervariable region of secukinumab determined by X-ray analysis, another preferred heavy chain framework is FR1-x (amino acids 1-25 of SEQ ID NO: 8), FR2-x (amino acids of SEQ ID NO: 8) in order. 36-49), FR3-x (amino acids 61-95 of SEQ ID NO: 8) and FR4 (amino acids 119-127 of SEQ ID NO: 8). Similarly, the light chain framework consists of FR1 ′ (amino acids 1 to 23 of SEQ ID NO: 10), FR2 ′ (amino acids 36 to 50 of SEQ ID NO: 10), FR3 ′ (amino acids 58 to 89 of SEQ ID NO: 10) in order, and It consists of the FR4 ′ (amino acids 99 to 109 of SEQ ID NO: 10) region.

  In one embodiment, the IL-17 antibody or antigen-binding fragment thereof, eg secukinumab, comprises at least: a) a variable domain comprising in turn the hypervariable regions CDR1, CDR2 and CDR3 of the human heavy chain and a constant portion or fragment thereof; The CDR1 has the amino acid sequence of SEQ ID NO: 1, the CDR2 has the amino acid sequence of SEQ ID NO: 2, the CDR3 has the amino acid sequence of SEQ ID NO: 3, and b) a human Comprising a variable domain comprising a light chain hypervariable region CDR1 ′, CDR2 ′ and CDR3 ′ in order and a constant portion or fragment thereof, said CDR1 ′ having the amino acid sequence of SEQ ID NO: 4, said CDR2 ′ being SEQ ID NO: 5 The CDR 3 ′ has the amino acid sequence of SEQ ID NO: 6, Roblin light chain or fragment thereof, is selected from a human anti-IL-17 antibody comprising a.

  In one embodiment, the IL-17 antibody or antigen-binding fragment thereof, eg secukinumab, comprises: a) the hypervariable regions CDR1, CDR2 and CDR3 in order, said CDR1 having the amino acid sequence of SEQ ID NO: 1, wherein said CDR2 is A first domain having the amino acid sequence of SEQ ID NO: 2, wherein the CDR3 has the amino acid sequence of SEQ ID NO: 3; and b) hypervariable regions CDR1 ′, CDR2 ′ and CDR3 ′ in order, the CDR1 ′ A second domain having the amino acid sequence of SEQ ID NO: 4, wherein the CDR2 'has an amino acid sequence of SEQ ID NO: 5, and the CDR3' has an amino acid sequence of SEQ ID NO: 6; and c) of the first domain N-terminal tip and C-terminal tip of second domain, or C-terminal tip of first domain and N-terminal tip of second domain Peptide linkers that crab bonded, are selected from a single chain binding molecule which comprises an antigen binding site comprising a.

Alternatively, an IL-17 antibody or antigen-binding fragment thereof, eg, secukinumab, for use in the disclosed methods can comprise a derivative of a molecule shown herein by sequence (eg, a PEGylated version of secukinumab). Alternatively, IL-17 antibody or antigen-binding fragment thereof for use in the methods disclosed, for example, V _H or V _L domains of secukinumab is, V _H or V _L domains shown herein (e.g., SEQ ID NO: 8 and Or a V _H or V _L domain that is substantially identical to that shown in FIG. The human IL-17 antibody disclosed herein comprises a heavy chain that is substantially identical to that shown in SEQ ID NO: 15 and / or a light chain that is substantially identical to that shown in SEQ ID NO: 14. Can do. The human IL-17 antibody disclosed herein can comprise a heavy chain comprising SEQ ID NO: 15 and a light chain comprising SEQ ID NO: 14. The human IL-17 antibody disclosed herein includes: a) one heavy chain comprising a variable domain having an amino acid sequence substantially identical to that shown in SEQ ID NO: 8 and a constant portion of a human heavy chain; b) a light chain comprising a variable domain having an amino acid sequence substantially identical to that shown in SEQ ID NO: 10 and a constant portion of a human light chain. Alternatively, an IL-17 antibody or antigen-binding fragment thereof, eg, secukinumab, for use in the disclosed methods may be an amino acid sequence variant of the reference molecule shown herein. In all such cases of derivatives and variants, the IL-17 antibody or antigen-binding fragment thereof, eg, secukinumab, exhibits an activity of about 1 nM (= 30 ng / ml) human IL-17 of about 50 nM or less of the molecule. About 20 nM or less, about 10 nM or less, about 5 nM or less, about 2 nM or less, more preferably about 1 nM or less, and the inhibitory activity is hu--in human skin fibroblasts. Measured by IL-6 production induced by IL-17.

Inhibition of IL-17 binding to its receptor can be conveniently tested in a variety of assays, including those described in WO2006 / 013107. The term “to the same extent” means that the reference and derivative molecules statistically exhibit essentially the same IL-17 inhibitory activity in one of the assays referred to herein (WO 2006/013107). See Example 1). For example, an IL-17 antibody or antigen-binding fragment thereof disclosed herein is induced by human IL-17 in human dermal fibroblasts when assayed as described in Example 1 of WO2006 / 013107. With respect to IL-6 production, inhibition of human IL-17 is less than about 10 nM, more preferably about 9, 8, 7, 6, 5, 4, 3, 2 or about 1 nM of the IC ₅₀ of the corresponding reference molecule. Preferably having an IC ₅₀ that is preferably substantially the same. Alternatively, the assay used is for IL-17 binding by a soluble IL-17 receptor (eg, the human IL-17R / Fc construct of Example 1 of WO2006 / 013107) and the IL-17 antibody or antigen-binding fragment thereof of the present disclosure. It may be an assay for competitive inhibition.

The present disclosure, for example mutation, for example by site-directed mutagenesis of the corresponding DNA sequence, the amino acid residue of the V _H or V _L domains as compared to V _H or V _L domains are shown in SEQ ID NO: 8 and SEQ ID NO: 10 Also included are IL-17 antibodies or antigen-binding fragments thereof, such as secukinumab, in which only one or more of the groups are changed, typically only a few (eg, 1-10). The present disclosure includes DNA sequences that encode such altered IL-17 antibodies.

  The present disclosure relates to IL-17 antibodies or antigen-binding fragments thereof having binding specificity to human IL-17, such as IL-17 antibodies capable of inhibiting the binding of IL-17 to secukinumab, particularly its receptor. , And 1 nM (= 30 ng / ml) of human IL-17 at a concentration of about 50 nM or less, about 20 nM or less, about 10 nM or less, about 5 nM or less, about 2 nM or less, more preferably about 1 nM or less of the molecule. Also included are IL-17 antibodies that are capable of 50% inhibition (the inhibitory activity is measured by IL-6 production induced by hu-IL-17 in human skin fibroblasts).

In some embodiments, an IL-17 antibody, such as secukinumab, binds to an epitope of mature human IL-17, including Leu74, Tyr85, His86, Met87, Asn88, Val124, Thr125, Pro126, Ile127, Val128, His129. In some embodiments, an IL-17 antibody, such as secukinumab, binds to an epitope of mature human IL-17, including Tyr43, Tyr44, Arg46, Ala79, Asp80. In some embodiments, an IL-17 antibody, such as secukinumab, binds to an epitope of an IL-17 homodimer having two mature human IL-17 chains, said epitope being on one chain Leu74, Tyr85, His86, Contains Met87, Asn88, Val124, Thr125, Pro126, Ile127, Val128, His129, and Tyr43, Tyr44, Arg46, Ala79, Asp80 on the other chain. The residue numbering scheme used to define these epitopes is based on residue 1 being the first amino acid of the mature protein (ie, lacking the 23 amino acid N-terminal signal peptide and starting with glycine) 17A). The sequence of immature IL-17A is shown in Swiss-Prot accession Q16552. In some embodiments, IL-17 antibody has a _{K D} of about 100~200pM for example when measured by Biacore (TM). In some embodiments, the IL-17 antibody has an IC ₅₀ of about 0.4 nM for in vitro neutralization of a biological activity of about 0.67 nM human IL-17A. In some embodiments, the absolute bioavailability of an IL-17 antibody administered subcutaneously (sc) has a range of about 60 to about 80%, such as about 76%. In some embodiments, the IL-17 antibody, eg, secukinumab, has an elimination half-life of about 4 weeks (eg, about 23 to about 35 days, about 23 to about 30 days, eg, about 30 days). In some embodiments, the IL-17 antibody, eg, secukinumab, has a T _{max of} about 7-8 days.

Particularly preferred IL-17 antibodies or antigen-binding fragments thereof, such as secukinumab, for use in the disclosed methods, uses, kits, etc., are human antibodies, particularly WO 2006/013107 (incorporated herein by reference in their entirety, US 7,807,155) secukinumab as described in Examples 1 and 2. Sekukinumab is an IgG1 / kappa isotype recombinant high affinity fully human monoclonal anti-human interleukin-17A (IL-17A, IL-17) that is currently in clinical trials for the treatment of immune-mediated inflammatory conditions It is an antibody. In secukinumab (see, for example, WO2006 / 013,107 and WO2007 / one hundred seventeen thousand seven hundred and forty-nine) have very high affinity for IL-17, i.e., _{K D} of about 100～200PM (e.g., as measured by Biacore (TM)) There is an IC ₅₀ for in vitro neutralization of the biological activity of about 0.67 nM human IL-17A is about 0.4 nM. Thus, secukinumab inhibits antigen at a molar ratio of about 1: 1. This high binding affinity makes secukinumab antibodies particularly suitable for therapeutic applications. In addition, secukinumab is determined to have a very long half-life, i.e. about 4 weeks, which allows for long-term dosing intervals and is a special property when treating chronic lifetime disorders such as psoriasis.

The present disclosure generally relates to a medicament comprising a container having a headspace having less than about 12% oxygen in the headspace, and a liquid composition disposed in the container. A product is provided and the liquid composition comprises the IL-17 antibody or antigen-binding fragment thereof, such as secukinumab.

Containers The pharmaceutical products of the present disclosure use primary packaging, or containers, to store, transport and maintain the disclosed liquid compositions. Pharmaceutically acceptable containers for use as part of the disclosed pharmaceutical product include syringes (eg, available from Beckton Dickinson, Nuova Ompi, etc.), vials with stoppers, cartridges, auto-injectors, Patch pumps and infusion pens are included.

Headspace Oxygen We have determined that the stability of the IL-17 antibody or antigen-binding fragment thereof (e.g., secukinumab) in the disclosed liquid composition can cause oxygen in the container headspace of the pharmaceutical product to pass through an inert gas ( For example, argon, helium, nitrogen), preferably N ₂ , was determined to be enhanced by the inclusion of certain stabilizers (eg, methionine) while simultaneously replacing N ₂ . Specifically, a pharmaceutical product that has been purged of oxygen, as measured by, for example, SEC and RP-HPLC, i.e., having a container with less than about 12% oxygen in the headspace, is relative to an unpurged product. The present inventors have determined that they have improved stability.

  Modification of the oxygen content in the headspace using a purge (eg, a nitrogen purge) can be achieved during the filling stage and / or plugging stage (or both). Purge (eg, nitrogen purge) can be accomplished by actively introducing an inert gas (eg, using a needle) or while plugging.

  In some embodiments, the oxygen content in the headspace is less than about 12% (eg, less than about 10%, less than about 8%, less than about 6%, etc.). In some embodiments, the oxygen content in the headspace is less than about 6%. The oxygen content in the headspace can be monitored by laser light absorption spectroscopy or fluorescence quenching or gas chromatography. It will be appreciated that the oxygen content in the headspace of a given container may increase over time, for example due to leakage. Thus, as used herein, the phrase “oxygen content in the headspace” refers to the initial level of oxygen in the headspace of the container immediately after the product is sealed (eg, plugged).

Liquid composition The liquid composition of the present disclosure comprises at least one of the IL-17 antibodies or antigen-binding fragments thereof (eg, secukinumab) as described above, as well as at least one additional excipient, eg, a buffer, a surfactant. And stabilizer (s). In some embodiments, the liquid composition includes at least two additional excipients, such as a buffer and a stabilizer. In some embodiments, the liquid composition includes a buffer, at least one stabilizer, and a surfactant.

  In general, pharmaceutical compositions are formulated with excipients that are compatible with the intended route of administration (eg, oral compositions generally include an inert diluent or an edible carrier). Examples of routes of administration include parenteral (eg, intravenous), intradermal, subcutaneous, oral (eg, oral or inhalation), transdermal (topical), transmucosal and rectal. The liquid antibody composition of this disclosure is suitable for parenteral administration, such as intravenous, intramuscular, intraperitoneal or subcutaneous injection, and particularly suitable for subcutaneous injection.

  In some embodiments, the liquid composition of the present disclosure has a purity of at least about 86% by RP-HPLC after 6 months storage at 2-8 ° C., and RP − after 6 months storage at 25 ° C./60% RH. Maintain at least about 76% purity by HPLC (preferably at least about 76%) and / or at least about 60% purity by RP-HPLC after 6 months storage at 30 ° C./75% RH. In some embodiments, the liquid compositions of the present disclosure maintain a purity of at least about 84% by RP-HPLC after storage at 2-8 ° C. for 24 months.

  In some embodiments, a liquid composition of the present disclosure is at least about 77% pure by CEX after 6 months storage at 2-8 ° C., and at least about CEX after 6 months storage at 25 ° C./60% RH Maintain 62% purity and / or at least about 50% purity by CEX after 6 months storage at 30 ° C./75% RH. In some embodiments, the liquid compositions of the present disclosure maintain a purity of at least about 73% by CEX after 24 months storage at 2-8 ° C.

  In some embodiments, the liquid composition of the present disclosure is at least about 98% pure by SEC after 6 months storage at 2-8 ° C., and at least about SEC by SEC after 6 months storage at 25 ° C./60% RH. Maintain a purity of 96% and / or a purity of at least about 94% by SEC after 6 months storage at 30 ° C./75% RH. In some embodiments, the liquid compositions of the present disclosure maintain a purity of at least about 97% by SEC after 24 months storage at 2-8 ° C.

  In some embodiments, a liquid composition of the present disclosure is at least about 97% pure by CE-SDS (non-reducing conditions) after 6 months storage at 2-8 ° C., 6 months at 25 ° C./60% RH. At least about 95% purity by CE-SDS (non-reducing conditions) after storage and / or at least about 94% by CE-SDS (non-reducing conditions) after storage for 6 months at 30 ° C./75% RH (preferably Maintaining a purity of at least about 92%). In some embodiments, a liquid composition of the present disclosure maintains a purity of at least about 97% by CE-SDS (non-reducing conditions) after storage at 2-8 ° C. for 24 months.

  In some embodiments, a liquid composition of the present disclosure has less than about 0.57% impurities by CE-SDS (reducing conditions) after 6 months storage at 2-8 ° C., 6% at 25 ° C./60% RH. Less than about 1.1% impurities by CE-SDS (reducing conditions) after storage for 6 months and / or less than about 1.9% by CE-SDS (reducing conditions) after 6 months storage at 30 ° C./75% RH Maintain the impurities. In some embodiments, the liquid compositions of the present disclosure maintain less than about 0.91% impurities by CE-SDS (non-reducing conditions) after storage at 2-8 ° C. for 24 months.

  In some embodiments, a liquid composition of the present disclosure has a relative biology of at least about 88% by inhibiting IL-6 release from C-20 / A4 chondrocytes after 24 months storage at 2-8 ° C. Activity, at least about 94% relative biological activity by inhibition of IL-6 release from chondrocytes after 6 months storage at 25 ° C./60% RH, and / or 6 months at 30 ° C./75% RH Maintain a relative biological activity of at least about 85% by inhibiting IL-6 release from chondrocytes after storage.

Antibody Concentration The IL-17 antibody or antigen-binding fragment thereof (eg, secukinumab) used in the disclosed liquid composition is described above. A preferred composition comprises secukinumab. We determined that antibody concentration had no significant effect on composition stability, at least within the range of about 25 mg / ml to about 150 mg / ml. Thus, in some embodiments, the antibody in the liquid composition is present at a concentration of at least 25 mg / ml (eg, from about 25 mg / ml to about 150 mg / ml). In some embodiments, the concentration of the antibody in the liquid composition is at a high concentration of at least about 25 mg / mL, at least about 50 mg / ml, at least about 75 mg / ml, at least about 100 mg / mL or at least about 150 mg / ml. is there. In some embodiments, the concentration of the antibody in the liquid composition is a high concentration from about 25 mg / mL to about 150 mg / mL. In one embodiment, the concentration of secukinumab in the liquid composition is about 25 mg / ml. In one embodiment, the concentration of secukinumab in the liquid composition is about 150 mg / ml.

Buffer and pH
Suitable buffering agents for the disclosed liquid compositions include, but are not limited to, gluconate buffer, histidine buffer, citrate buffer, phosphate [eg, sodium or potassium] buffer, succinate Acid salts [eg, sodium] buffer, acetate buffer, Tris buffer, glycine, arginine and combinations thereof are included. The inventors determined that there was no beneficial effect of succinate or acetate buffer on the stability of the liquid composition of secukinumab. Citrate buffer was evaluated as beneficial in the composition with respect to SEC, CEX acid degradation products and RP-HPLC aggregation products. Overall, histidine buffer has shown advantages in agglomeration and degradation products with SEC, CEX acid and RP-B. Thus, histidine buffer is a preferred buffer for the disclosed stable liquid composition of secukinumab.

  Histidine buffer (e.g., about 5 mM to about 50 mM, e.g., about 20 mM to about 50 mM, about 5 mM, about 10 mM, about 15 mM, about 20 mM, about 25 mM, about 30 mM, about 35 mM, about 40 mM, about 45 mM, about 50 mM Concentration) is particularly beneficial. In one embodiment, the stable liquid composition comprises about 20 mM to about 50 mM histidine buffer. The pH of the liquid composition may be in the range of 4.0 to 8.0, such as a pH in the range of about 5.5 to about 7.4, such as about 5.2 to about 6.2, about 5 .2 to about 5.8, for example, about 5.2, about 5.3, about 5.4, about 5.5, about 5.6, about 5.7, about 5.8, about 5.9, About 6, about 6.2, about 6.4, about 6.6, about 6.7, about 6.8, about 6.9, about 7.0, about 7.1, about 7.2, about 7 .3, about 7.4 is common. By increasing the pH from 5.2 to 5.8, a positive trend of stability is observed (SEC-AP, DLS, SEC-DP, ALP-DP, CEX base, RP-HPLC) The inventors determined. Overall testing showed that the ideal composition pH of the disclosed liquid composition was 5.8. Thus, in one embodiment, the stable liquid antibody composition has a pH of about 5.8.

Surfactants Suitable surfactants for the disclosed liquid composition include, but are not limited to, nonionic surfactants, ionic surfactants, zwitterionic surfactants and combinations thereof. Common surfactants for the present invention include, but are not limited to, sorbitan fatty acid esters (eg, sorbitan monocaprylate, sorbitan monolaurate, sorbitan monopalmitate), sorbitan trioleate, glycerin fatty acid esters (eg, Glycerin monocaprylate, glycerin monomyristate, glycerin monostearate), polyglycerin fatty acid ester (eg decaglyceryl monostearate, decaglyceryl distearate, decaglyceryl monolinoleate), polyoxyethylene sorbitan fatty acid ester (eg , Polyoxyethylene sorbitan monolaurate, polyoxyethylene sorbitan monooleate, polyoxyethylene sorbitan monostearate, polyoxyethylene sorbitan monopal Tate, polyoxyethylene sorbitan trioleate, polyoxyethylene sorbitan tristearate), polyoxyethylene sorbitol fatty acid ester (for example, polyoxyethylene sorbitol tetrastearate, polyoxyethylene sorbitol tetraoleate), polyoxyethylene glycerin fatty acid ester (For example, polyoxyethylene glyceryl monostearate), polyethylene glycol fatty acid ester (for example, polyethylene glycol distearate), polyoxyethylene alkyl ether (for example, polyoxyethylene lauryl ether), polyoxyethylene polyoxypropylene alkyl ether ( For example, polyoxyethylene polyoxypropylene glycol, polyoxyethylene polyoxy Propylene propyl ether, polyoxyethylene polyoxypropylene cetyl ether), polyoxyethylene alkylphenyl ether (eg, polyoxyethylene nonylphenyl ether), polyoxyethylene hydrogenated castor oil (eg, polyoxyethylene castor oil, polyoxyethylene hydrogenated castor) Oil), polyoxyethylene beeswax derivatives (eg, polyoxyethylene sorbitol beeswax), polyoxyethylene lanolin derivatives (eg, polyoxyethylene lanolin) and polyoxyethylene fatty acid amides (eg, polyoxyethylene stearamide); C10-C18 alkyl sulfate (eg, sodium cetyl sulfate, sodium lauryl sulfate, sodium oleyl sulfate), an average of 2-4 moles of ethylene oxide units added. Reoxyethylene C10-C18 alkyl ether sulfate (eg, polyoxyethylene sodium lauryl sulfate) and C1-C18 alkyl sulfosuccinate (eg, lauryl sulfosuccinate sodium ester); and natural surfactants such as lecithin, glyceroline Lipids, sphingophospholipids (eg, sphingomyelin) and sucrose esters of C12 to C18 fatty acids are included. The composition can include one or more of these surfactants. Preferred surfactants are poloxamers (eg poloxamer 188) or polyoxyethylene sorbitan fatty acid esters such as polysorbate 20, 40, 60 or 80. Polysorbate 80 (Tween 80) (eg, about 0.01% to about 0.1% (w / v), eg, about 0.01% to about 0.04% (w / v), eg, about 0.01 %, About 0.02%, about 0.04%, about 0.06%, about 0.08%, about 0.1% concentration) are particularly useful. In one embodiment, the stable liquid composition comprises about 0.02% (w / v) polysorbate 80. In one embodiment, the stable liquid composition comprises about 0.02% (w / v) polysorbate 20.

  We have determined that liquid compositions lacking surfactants have a significant increase in turbidity, as well as an increase in the amount of visible particles. However, with the exception of increases in ALP-DP and RP, no benefit of poloxamer 188 was detected compared to polysorbates 20 and 80. Polysorbates 20 and 80 showed equivalent effectiveness in preventing turbidity, sub-visible and visible particle build-up. Polysorbates 20 and 80 are therefore preferred surfactants for use in the disclosed stable liquid compositions.

Stabilizers Stabilizers help prevent protein oxidation and aggregation in liquid pharmaceutical compositions that have a shorter shelf life due to the tendency of the protein to oxidize and / or aggregate in aqueous solutions. A variety of analytical methods can be used to assess the stability of a given composition, for example, assaying the level of oxidation products (pre-main peaks) with the liquid compositions disclosed herein. RP-HPLC can be used to do this, and SEC can be used to assay aggregation levels with the liquid compositions disclosed herein.

  Stabilizers suitable for use in the disclosed liquid compositions include ionic and non-ionic stabilizers (and combinations thereof) such as sugars, glycine, sodium chloride, arginine, EDTA, sodium ascorbate, cysteine , Sodium bisulfate, sodium citrate, methionine and benzyl alcohol. In some embodiments, the liquid pharmaceutical composition contains at least one stabilizer from a first group (eg, sugar [eg, trehalose, mannitol], amino acids [eg, glycine, arginine] and sodium chloride). To do. In some embodiments, the liquid pharmaceutical composition contains at least one stabilizer from the second group (EDTA, sodium ascorbate, cysteine, sodium bisulfate, sodium citrate, methionine and benzyl alcohol). A second group of stabilizers tends to have antioxidant properties, which can reduce oxidation of residues with IL-17 antibodies. In a preferred embodiment, the liquid pharmaceutical composition contains two stabilizers, one from the first group and one from the second group.

  For the first group of stabilizers, nonionic stabilizers are preferred. Suitable nonionic stabilizers include monosaccharides, disaccharides and trisaccharides such as trehalose, raffinose, maltose, sorbitol or mannitol. The sugar may be a sugar alcohol or an amino sugar. The concentration of the first group of stabilizers is about 175 mM to about 350 mM, such as about 200 mM to about 300 mM, such as about 250 mM to about 270 mM, such as about 180 to about 300 mM, about 200 mM to about 225 mM, about 175 mM, It may be about 180 mM, about 185 mM, about 190 mM, about 195 mM, about 200 mM, about 225 mM, about 250 mM, about 270 mM, 275 mM, about 300 mM. Mannitol at a concentration of about 200 mM to about 300 mM (eg, about 250 mM to about 270 mM), trehalose at a concentration of about 180 mM to about 300 mM (eg, about 200 mM to about 225 mM), sodium chloride at a concentration of about 130 mM to about 150 mM, about Particularly useful are arginine at a concentration of 160 mM, glycine at a concentration of about 270 mM.

  Although we determined that glycine as a stabilizer (first group) was slightly advantageous with respect to SEC-AP and DLS, an increase in almost all degradation products was observed. NaCl as a stabilizer (first group) led to degradation by SEC and CEX basic mutants and an increase in aggregated products. Trehalose and mannitol acted as equivalent beneficial stabilizers and were confirmed in almost all analyses, but mannitol had a slightly less effective effect (SEC-AP, DLS) in addition to trehalose and a lower water solubility. Indicated. Trehalose is therefore a first group of preferred stabilizers because of its positive impact on degradation products. In one embodiment, the liquid composition comprises about 200 mM to about 225 mM trehalose. In one embodiment, the liquid composition comprises about 200 mM trehalose. In one embodiment, the liquid composition comprises about 225 mM trehalose.

  We determined that there was a significant effect of the second group on the stability of the liquid composition of secukinumab. Our experiments show that the non-use of the second group of stabilizers is inferior compared to the composition containing the second group of stabilizers (SEC-AP, DLS, turbidity, RP-B ) Tetrasodium EDTA and cysteine showed increased aggregation and degradation products in each assay. The addition of cysteine as a second group of stabilizers resulted in a cloudy composition after freeze-thaw stress and precipitation within 4 weeks of storage at 40 ° C. However, the inventors have determined that methionine is advantageous in all compositions for analysis. Therefore, for the second group of stabilizers, methionine that also has antioxidant properties is preferred. The concentration of the second group of stabilizers (eg, methionine) is at least about 2.5 mM, such as about 2.5 to about 20 mM, such as at least about 2.5 mM, at least about 5 mM, at least about 10 mM, or at least about 20 mM ( For example, it may be about 2.5 mM, about 5 mM, about 10 mM, or about 20 mM). In a preferred embodiment, the liquid pharmaceutical composition contains at least one stabilizer from the first group and methionine. In some embodiments, the disclosed liquid composition comprises about 5 mM methionine.

Other Excipients The liquid antibody composition of the present disclosure may be further excipients such as additional buffers, salts (eg, sodium chloride, sodium succinate, sodium sulfate, potassium chloride, magnesium chloride, magnesium sulfate and chloride). Calcium), additional stabilizers, tonicity modifiers (eg, salts and amino acids [eg, proline, alanine, L-arginine, asparagine, L-aspartic acid, glycine, serine, lysine and histidine]), glycerol, albumin , Alcohols, preservatives, additional surfactants, antioxidants, and the like. A complete discussion of such additional pharmaceutical ingredients is available in Gennaro (2000) Remington: The Science and Practice of Pharmacy. 20th edition, ISBN: 0683306472.

Additional Active Agents The pharmaceutical products and stable liquid compositions of the present disclosure include one or more other active agents (eg, psoriasis agents, psoriatic agents) in addition to the IL-17 antibody or antigen-binding fragment thereof, eg, secukinumab. Arthritis drug, ankylosing spondylitis drug, rheumatoid arthritis drug). Such additional factors and / or to produce a synergistic effect with the IL-17 antibody or antigen-binding fragment thereof, or to minimize side effects due to the IL-17 antibody or antigen-binding fragment thereof, eg secukinumab Alternatively, a drug may be included in the pharmaceutical composition.

  Examples of psoriatic agents that may be formulated with the disclosed IL-17 antibodies such as secukinumab include cyclosporine, methotrexate, mycophenolate mofetil, mycophenolate, sulfasalazine, 6-thioguanine, fumarate (eg, , Dimethyl fumarate and fumaric acid esters), azathioprine, corticosteroids, leflunomide, tacrolimus, T cell blockers (eg Amive® (alephacept) and Raptiva® (efalizumab), tumor necrosis factor alpha (TNF) -Alpha) blockers (e.g. Enbrel (R) (etanercept), Humira (R) (adalimumab), Remicade (R) (infliximab) and Simoni R) (golimumab)) and interleukin 12/23 blockers (e.g. Stelara (TM) (Usutekinumabu) include Tasoshichinibu and Buriakinumabu.

  Additional psoriasis agents that may be formulated with the disclosed IL-17 antibodies such as secukinumab for the treatment of psoriasis include apremilast, mometazom, voclosporin, ketoconazole, Neuroskin Forte, recombinant human interleukin-10, Voclosporine, MK-3222, tofacitinib, VX-765, MED-I545, fluphenazine decanoate, acetominofun, bimosia moss cream, doxycycline, vancomycin, AbGn168, vitamin D3, RO5310074, fludarabine calcipotriol and hydrocortisone (LE80) Focusria (monovalent MF59 adjuvanted vaccine, tgAAC94 gene therapy vector, capsaicin, Psirelax, AB -874 (anti-IL-12), IDEC-114, MEDI-522, LE29102, BMS587101, CD2027, CRx-191, 8-methoxypsoralen or 5-methoxypsoralen, Bicillin LA, LY2525623, INCB018424, LY2439821, CEP-701, CC-10004, Sertolizumab (CZP), GW7866034 (pazopanib), doxycycline cuminoid C3 complex, NYC0462, RG3421, hOKT3 gamma1 (Ala-Ala), BT061, teplizumab, chondroitin sulfate, CNTO1240, IL Monoclonal antibodies against IL-23p40 subunit, BMS-582949, MK0873, MEDI-507, 518101, ABT-874, AMG827, AN2728, AMG714, AMG139, PTH (1-34), U0267 foam, CNTO1275, QRX-101, CNTO1959, LEO22811, Imiquimod, CTLA4Ig, Algae Dunaliella Bardawil, Dunaliella Bardawil, , Ranibizumab, Zidovudine CDP870 (Certolizumab Pegor), Onercept (r-hTBP-1), ACT-128800, 4,4-dimethyl-benzoiso-2H-selenazine, CRx-191, CRx-197, doxercalciferol , LAS41004, WBI-1001, tacrolimus, RAD001, rapamycin, rosiglitazone, pioglitazone, ABT-874, aminopte Emissions, AN2728, CD2027, ACT-128800, mometasone furoate, CT327, clobetasol + LCD, BTT1023, E6201, topical vitamin B12, IP10. C8, BFH772, LEO22811, fluphenazine, MM-093, Clobex, SCH527123, CF101, SRT2104, BIRT2584, CC10004, tetrathiomolybdate, CP-690,550, U0267, ASP015K, VB-201, acitretin (U0279) , RWJ-445380, clobetasol propionate, botulinum toxin type A, alefacept, erlotinib, BCT194, roflumilast, CNTO1275, halobetasol, ILV-094, CTA018 cream, COL-121, MEDI-507, AEB071.

  Additional psoriasis agents that may be formulated with the disclosed IL-17 antibodies such as secukinumab include IL-6 antagonist, CD20 antagonist, CTLA4 antagonist, IL-17 antagonist, IL-8 antagonist, IL-21 Antagonists, IL-22 antagonists, VGEF antagonists, CXCL antagonists, MMP antagonists, defensin antagonists, IL-1beta antagonists and IL-23 antagonists (eg, receptor decoys, antagonist antibodies, etc.) are included. Preferred psoriasis agents that may be formulated with secukinumab are DMARDs (eg MTX and cyclosporine), IL-12 / -23 antagonists (eg ustekinumab), CTLA-4 antagonists (eg CTLA4-Ig) and TNF An alpha antagonist.

  In a broad sense, rheumatoid arthritis drugs, psoriatic arthritis drugs and ankylosing spondylitis drugs that may be formulated with the disclosed IL-17 antibodies such as secukinumab include immunosuppressants, DMARDs, pain management, among others. It may be a drug, a steroid, a non-steroidal anti-inflammatory drug (NSAID), a cytokine antagonist, a bone anabolic agent, a bone anti-resorptive agent and combinations thereof. Representative drugs include cyclosporine, retinoids, corticosteroids, propionic acid derivatives, acetic acid derivatives, enolic acid derivatives, fenamic acid derivatives, Cox-2 inhibitors, lumiracoxib, magnesium ibuprofen choline salicylate, fenoprofen, salsalate, difunisal , Tolmetine, ketoprofen, flurbiprofen, oxaprozin, indomethacin, sulindac, etodolac, ketorolac, nabumetone, naproxen, valdecoxib, etoroxib, MK0966; lofecoxib, acetminophen, celecoxib, diclofenac, tramadol, piroxicam Lornoxicam, isoxicam, mephanamic acid, meclofenamic acid, flufenamic acid, tolfenam Valdecoxib, parecoxib, etodolac, indomethacin, aspirin, ibuprofen, phylocoxib, methotrexate (MTX), antimalarials (eg, hydroxychloroquine and chloroquine), sulfasalazine, leflunomide, azathioprine, cyclosporine, gold salt, minocycline, cyclophosphamide, D-penicillamine, minocycline, auranofin, tacrolimus, myoclysin, chlorambucil, TNF alpha antagonists (eg TNF alpha antagonists or TNF alpha receptor antagonists) such as adalimumab (Humira®), etanercept (Enbrel® )), Infliximab (Remicade®; TA-650), Trizumab Pegor (Cimzia®; CDP870), Golimumab (Simponi®; CNTO148), Anakinra (Kineret®), Rituximab (Rituxan®; MabThera®), Abatacept (Orencia®), tocilizumab (RoActemra / Actemra®), integrin antagonist (TYSABRI® (natalizumab)), IL-1 antagonist (ACZ885 (Ilaris)), anakinra (Kineret®) )), CD4 antagonist, IL-23 antagonist, IL-20 antagonist, IL-6 antagonist, BLyS antagonist (eg, atacicept, Be nlysta <(R)> / LymphoStat-B <(R)> (berimumab)), p38 inhibitor, CD20 antagonist (Ocrelizumab, ofatumumab (Arzerra <(R)>)), interferon-gamma antagonist (Fontlizumab), prednisolone, prednisone, Dexamethasone, Cortisol, Cortisone, Hydrocortisone, Methylprednisolone, Betamethasone, Triamcinolone, Beclometazome, Fludrocotisone, Deoxycorticosterone, Aldosterone, SB-681323, Rob 803, AZD5672, AD 452, SMP 114, HZT-541 , Doxycycline, vancomycin, CRx-102, AMG108, pioglitazone, SBI- 87, SCIO-469, Cura-100, Oncoxin + bisulfide, TwHF, PF-0471327, AZD5672, methoxalene, ARRY-438162, vitamin D-ergocalciferol, milnacipran, paclitaxel, GW406381, rosiglitazone, SC12267 (4SC- 101); LY2439821, BTT-1023, ERB-041, ERB-041, KB003, CF101, ADL5859, MP-435, ILV-094, GSK706769, GW856553, ASK8007, MOR103, HE3286, CP-690,550 (Tasocitinib) REGN88 (SAR153191), TRU-015, BMS-582949, SBI-087, LY2127399, E- 51S-551, H-551, GSK3152314A, RWJ-445380, tacrolimus (Prograf (registered trademark)), RAD001, rapamun, rapamycin, fostatinib, fentanyl, XOMA052, CNTO136, JNJ3851810IS, N-ima1038AT , Folic acid, Folate, TNFa quinoid, MM-093, Type II collagen, VX-509, AMG 827 70, Masitinib (AB1010), LY2127399, Cyclosporine, SB-681233, MK0663, NNC 0151-0000-0000, ATN-103 , CCX 354-C, CAM 3001, LX 3305, Setro Relix, MDX-1342, TMI-005, MK0873, DP870, tranilast, CF101, mycophenolic acid (and its ester), VX-702, GLPG0259, SB-681323, BG9924, ART621, LX3305, T-614, fosamatinib disodium (R935788), CCI-779, ARRY-371797, CDP6038, AMG719, BMS-582949, GW856553, rosiglitazone, CH-4051, CE-224,535, GSK1827771, GW274150, BG9924, PLX3397, TAK-783, INCB028050, LY2127399, LY3009104, R788on Rosuvastatin, PRO283698, AMG 714, MTRX1011A, Rabilok, MEDI-522, MK0663, STA 5326 mesylic acid, CE-224,535, AMG108, BG00012 (BG-12; Biogen), ramipril, VX-702, CRx-102, LY2189102, SBI-087, SB-681323, CDP870, milnacipran, PD0360324, PH-79804, AK106-001616, PG-760564, PLA-695, MK0812, ALD518, cobiprostone, somatropin, tgAAC94 gene therapy vector, MK0359, GW856553, ezomeprazole, everolimus, trastuzumab Anabolic and bone anti-resorptive drugs (eg PTH, bisphosphonates (eg zoledronic acid), JAK1 and JAK2 inhibitors, pan-J AK inhibitors, such as tetracyclic pyridone 6 (P6), 325, PF-95980, sclerostin antagonists (eg, WO09047356, WO2000 / 32773, WO2006102070, US Patent Application Publication No. 200802227138, US Patent Application Publication No. 20120223535, United States) Patent Application Publication No. 20030290441, WO2005003158, WO2009039175, WO20090907471, WO03106657, WO2006119062, WO08115732, WO2005 / 014650, WO2005 / 003158, WO2006 / 119107, WO2008 / 061013, WO2008 / 133724, WO2008 / 115724 No. 7879322, US Pat. No. 7,744,874, the contents of which are hereby incorporated by reference in their entirety [preferred anti-sclerostin antibodies for use in the disclosed methods, pharmaceutical compositions, kits and applications and Its antigen-binding portion is found in WO09047356 (equivalent to US Pat. No. 7,879,322), WO06119107 (equivalent to US Pat. No. 7,872,106 and US Pat. No. 7,592,429) and WO08115732 (equivalent to US Pat. No. 7,744,874)], denosumab, IL-6 antagonist, CD20 antagonist, CTLA4 antagonist, IL-8 antagonist, IL-21 antagonist, IL-22 antagonist, integrin antagonist (Tysarbri®) (Natalizumab)), VGEF antagonists, CXCL antagonists, MMP antagonists, defensin antagonists, IL-1 antagonists (including IL-1beta antagonists) and IL-23 antagonists (eg, receptor decoys, antagonist antibodies, etc.). Preferred rheumatoid arthritis agents that may be formulated with the disclosed IL-17 antibodies such as secukinumab are DMARDs such as methotrexate and TNF alpha antagonists. Preferred ankylosing spondylitis agents that may be formulated with the disclosed IL-17 antibodies such as secukinumab are NSAIDs, DMARDs such as sulfasalazine, and TNF alpha antagonists. Preferred psoriatic arthritis agents that may be formulated with the disclosed IL-17 antibodies such as secukinumab are DMARDs such as cyclosporine, CTLA-4 blockers (eg CLTA4-Ig), alefacept and TNF alpha antagonists .

  One skilled in the art can determine appropriate dosages of the above agents for co-composition with the disclosed IL-17 antibodies, such as secukinumab.

  As used herein, from about 20 mg / ml to about 175 mg / ml (eg, about 25 mg / ml to about 150 mg / ml) of an IL-17 antibody or antigen-binding fragment thereof (eg, secukinumab) disclosed herein, About 10 mM to about 30 mM buffer (eg, histidine) pH 5.2 to about 6.0, about 200 mM to about 225 mM stabilizer (eg, trehalose), about 0.02% surfactant (eg, polysorbate 80 ) And about 2.5 mM to about 20 mM methionine is disclosed.

  In some embodiments, the concentration of methionine in the liquid pharmaceutical composition of the disclosed pharmaceutical is about 2.5 mM, about 5 mM, about 10 mM or about 20 mM, preferably about 5 mM. In some embodiments, the pH of the liquid pharmaceutical composition is about 5.8. In some embodiments, the secukinumab concentration of the disclosed compositions is about 25 mg / ml or about 150 mg / ml. In some embodiments, the liquid pharmaceutical composition comprises a buffer selected from the group consisting of a histidine buffer, a citrate buffer, an acetate buffer, and a succinate buffer. In some embodiments, the liquid pharmaceutical composition uses a histidine buffer at a concentration of about 20 mM. In some embodiments, the liquid pharmaceutical composition comprises a surfactant selected from polysorbate and poloxamer. In some embodiments, the liquid pharmaceutical composition further comprises a surfactant selected from polysorbate 80, polysorbate 20, and poloxamer 188. In some embodiments, the liquid pharmaceutical composition has a concentration of about 0.01% (w / v) to about 0.04% (w / v), preferably about 0.02% (w / v). Polysorbate 80 is included. In some embodiments, the liquid pharmaceutical composition comprises polysorbate 20 at a concentration of about 0.02% (w / v). In some embodiments, the liquid pharmaceutical composition comprises a stabilizer selected from the group consisting of mannitol, sodium chloride, trehalose, arginine HCl and glycine. In some embodiments, the liquid pharmaceutical composition comprises trehalose at a concentration of about 180 mM to about 300 mM, preferably about 200 mM or about 225 mM.

  As used herein, a container having a headspace in which the oxygen content is less than about 12%, and a liquid pharmaceutical composition disposed in the container, wherein the liquid pharmaceutical composition is about 5.2 to about 6.2. From about 20 mg / ml to about 175 mg / ml (eg, from about 25 mg / ml to about 150 mg / ml) of an IL-17 antibody or antigen-binding fragment thereof (eg, secukinumab) disclosed herein And a liquid pharmaceutical composition comprising about 2.5 to about 20 mM L-methionine and not reconstituted from the lyophilizate.

  In some embodiments, the concentration of methionine in the liquid pharmaceutical composition of the disclosed pharmaceutical is about 2.5 mM, about 5 mM, about 10 mM or about 20 mM, preferably about 5 mM. In some embodiments, the oxygen content in the headspace of the disclosed pharmaceutical product is less than about 10%, such as less than about 8%, preferably less than about 6%. In some embodiments, the liquid pharmaceutical composition of the disclosed pharmaceutical product has a pH of about 5.8. In some embodiments, the disclosed pharmaceutical product has a secukinumab concentration of about 25 mg / ml or about 150 mg / ml. In some embodiments, the liquid pharmaceutical composition of the disclosed pharmaceutical product further comprises a buffer selected from the group consisting of a histidine buffer, a citrate buffer, an acetate buffer, and a succinate buffer. In some embodiments, the liquid pharmaceutical composition of the disclosed pharmaceutical product uses a buffer at a concentration of about 10 mM to about 30 mM. In some embodiments, the liquid pharmaceutical composition of the disclosed pharmaceutical product uses a histidine buffer at a concentration of about 20 mM. In some embodiments, the liquid pharmaceutical composition of the disclosed pharmaceutical product further comprises a surfactant selected from polysorbate and poloxamer. In some embodiments, the liquid pharmaceutical composition of the disclosed pharmaceutical product further comprises a surfactant selected from polysorbate 80, polysorbate 20, and poloxamer 188. In some embodiments, the liquid pharmaceutical composition of the disclosed pharmaceutical product is about 0.01% (w / v) to about 0.04% (w / v), preferably about 0.02% (w It further comprises polysorbate 80 at a concentration of / v). In some embodiments, the liquid pharmaceutical composition of the disclosed pharmaceutical product further comprises polysorbate 20 at a concentration of about 0.02% (w / v). In some embodiments, the liquid pharmaceutical composition of the disclosed pharmaceutical product further comprises a stabilizer selected from the group consisting of mannitol, sodium chloride, trehalose, arginine HCl and glycine. In some embodiments, the liquid pharmaceutical composition of the disclosed pharmaceutical product further comprises trehalose at a concentration of about 180 mM to about 300 mM, preferably about 200 mM or about 225 mM. In some embodiments, the disclosed pharmaceutical product containers are cartridges, syringes, pens or vials.

  As used herein, a container having a headspace in which the oxygen content is less than about 6%, and a liquid pharmaceutical composition disposed in the container, wherein the liquid pharmaceutical composition is about 20 mg / ml to about 175 mg / ml. (Eg, about 25 mg / ml to about 150 mg / ml) of an IL-17 antibody disclosed herein or an antigen-binding fragment thereof (eg, secukinumab), about 10 mM to about 30 mM histidine pH 5.8, about 200 mM to Disclosed is a pharmaceutical product comprising a liquid pharmaceutical composition comprising about 225 mM trehalose, about 0.02% polysorbate 80, and about 2.5 mM to about 20 mM methionine that has not been reconstituted from the lyophilizate.

  In some embodiments, the pharmaceutical product comprises about 25 mg / ml secukinumab and about 225 mM trehalose. In some embodiments, the pharmaceutical product comprises about 150 mg / ml secukinumab and about 200 mM trehalose. In some embodiments, the disclosed pharmaceutical product containers are cartridges, syringes, pens or vials.

  In some embodiments, the pharmaceutical product has an amount of IL- sufficient to allow delivery of at least about 75 mg to about 300 mg of an IL-17 antagonist (eg, an IL-17 antibody, such as secukinumab) per unit dose. Has 17 antagonists. In some embodiments, the pharmaceutical product has an amount of an IL-17 antagonist (eg, an IL-17 antibody such as secukinumab) sufficient to allow delivery of at least about 10 mg / kg per unit dose. In some embodiments, the pharmaceutical product is formulated at a dosage that allows intravenous delivery of about 10 mg / kg of an IL-17 antagonist (eg, an IL-17 antibody, eg, secukinumab) per unit dose. In some embodiments, the pharmaceutical product is formulated at a dosage that allows subcutaneous delivery of at least about 75 mg to about 300 mg of an IL-17 antagonist (eg, IL-17 antibody, eg, secukinumab) per unit dose. .

Methods for Making Liquid Compositions and Pharmaceutical Products Methods for making the pharmaceutical products and liquid compositions of the present disclosure are also described herein. These methods help reduce oxidation of the disclosed IL-17 antibodies. Briefly, the liquid composition comprises a desired excipient (eg, a first group of stabilizers (eg, trehalose), a second group of stabilizers (methionine), a surfactant (eg, PS80), A buffer (eg, histidine)) and an IL-17 antibody or antigen-binding fragment thereof (eg, secukinumab) and a desired concentration (eg, about 25 to about 150 mg / ml secukinumab, about 20 mM histidine pH 5.8, about 200 mM to About 225 mM trehalose, about 0.02% polysorbate 80 and about 2.5 mM to about 20 mM methionine) and pH (eg, about pH 5.8). The liquid composition is then placed in a selected container (eg, vial, syringe, cartridge [eg, for autoinjector]). The oxygen content in the headspace is adjusted to the desired level (eg, less than about 12%, less than 10%, less than about 8%, less than about 6%, etc.), but this is done with a liquid composition in a container May be before filling the container with the liquid composition, or while plugging / sealing the container.

  As used herein, having a pH of about 5.2 to about 6.2 and disclosed herein from about 20 mg / ml to about 175 mg / ml (eg, from about 25 mg / ml to about 150 mg / ml). Preparing a liquid composition comprising an IL-17 antibody or antigen-binding fragment thereof (eg, secukinumab) and from about 2.5 mM to about 20 mM methionine; placing said liquid composition in a container having a headspace And a method for reducing oxidation of secukinumab comprising adjusting the oxygen content in the headspace to about 12% or less.

  In some embodiments of the disclosed method, the conditioning step c) is performed by purging the headspace using an inert gas. In some embodiments of the disclosed method, the inert gas is nitrogen or argon. In some embodiments of the disclosed methods, the concentration of methionine in the liquid composition is about 2.5 mM, about 5 mM, about 10 mM or about 20 mM, preferably about 5 mM. In some embodiments of the disclosed method, the oxygen content in the headspace is adjusted to less than about 10%, such as less than about 8%, preferably less than about 6%. In some embodiments of the disclosed method, the liquid composition has a pH of about 5.8. In some embodiments of the disclosed method, the concentration of secukinumab in the liquid composition is about 25 mg / ml or about 150 mg / ml. In some embodiments of the disclosed method, the container is a cartridge, syringe, pen or vial.

Methods of using pharmaceutical products and liquid compositions The disclosed pharmaceutical products and liquid compositions are used, for example, in patients with autoimmune diseases (eg, psoriasis, rheumatoid arthritis, ankylosing spondylitis, psoriatic arthritis, etc.) Used for treatment. Suitable dosages include, for example, the particular IL-17 antibody or antigen-binding fragment thereof used, such as secukinumab, host, mode of administration, and the nature and severity of the condition being treated, and the nature of the previous treatment the patient has received Of course, it depends on. Ultimately, the healthcare provider in charge will determine the amount of IL-17 antibody to treat each individual patient. In some embodiments, the healthcare provider can administer a low dose of IL-17 antibody and observe the patient's response. In other embodiments, the initial dose (s) of IL-17 antibody administered to the patient is high and then dosed downward until signs of recurrence appear. Larger doses of IL-17 antibody can be administered until the optimal therapeutic effect is obtained in the patient, and the dosage is generally not increased further.

  The timing of administration is generally measured from the first day of administration of the active compound (eg, secukinumab), also known as “baseline”. However, as shown in Table 2 below, different health care providers use different nomenclature.

  Of note, week 0 may be referred to as week 1 by some health care providers, and day 0 may be referred to as day 1 by some health care providers. Thus, different physicians may refer to the same dosing schedule, for example, if the dose is between Week 3 / Day 21, Week 3, Week 22 / Day 4, Week 21 / Day 21, Week 4 May be referred to as administered between eyes / day 22. For consistency, the first week of administration is referred to herein as week 0 and the first day of administration is referred to as day 1. However, those skilled in the art will appreciate that this nomenclature is only used for consistency and should not be construed as limiting, i.e., the physician may designate a particular week as "Week 1" or "Week 2". Weekly administration is the provision of weekly doses of IL-17 antibody or antigen-binding fragment thereof, such as secukinumab. As an example of nomenclature using the rules specified herein, five doses of secukinumab administered weekly are given during week 0 (eg around day 1), during week 1 (eg day 8). Around), 2 weeks (eg around day 15), 3 weeks (eg around day 22) and 4 weeks (eg around day 29). . As long as it is provided in the appropriate week, the dose need not be provided at the exact time, for example, the scheduled dose on day 29 is provided, for example, on days 24-34, eg, day 30 It is understood that this may be done.

  In some embodiments, the disclosed methods and uses include initials lasting 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15 or 16 weeks Use a regimen (sometimes called "introduction"). In some embodiments, the initial regimen uses dosing during the 0, 1, 2, and 3 week periods. In other embodiments, the initial regimen uses dosing during the 0, 1, 2, 3, 4, 8 and 12 week periods. In some embodiments, the initial regimen is some 150 mg to 300 mg (eg 1, 2, 3, 4, 5, 6, 7, preferably 4 or 5) of an IL-7 antibody, eg secukinumab. Administration of a dose, for example 150 mg or 300 mg of about 4 or 5 doses (preferably about 150 mg to about 300 mg of 5 doses). In further embodiments, the initial dose is delivered weekly, twice weekly, biweekly or monthly [every 4 weeks], preferably weekly. In some embodiments, IL-17 antibody, eg, 150 mg or 300 mg of secukinumab is administered by subcutaneous injection at the initial doses at 0, 1, 2, and 3 weeks.

  For maintenance regimens, the dose is monthly (also called “monthly” administration) (ie every 4 weeks, ie about every 28 days), every 2 months (ie every 8 weeks, ie around every 56 days). Alternatively, it can be provided every 3 months (ie every 12 weeks, ie approximately every 84 days). In some embodiments, the maintenance regimen begins after the 12th week. In some embodiments, the maintenance regimen begins after the third week. The first dose of the maintenance regimen is administered on the day usually measured from the final dose of the induction regimen. Thus, for example, if the final dose of the induction regimen is provided during week 12, the first dose as part of the monthly [every 4 week] maintenance regimen is delivered during week 16 and 2 The first dose as part of a monthly maintenance regimen will be delivered during week 20, the first dose as part of every 3 months maintenance regimen will be delivered during week 24, etc. . In some embodiments, the maintenance regimen comprises a dose of IL-17 antibody or antigen-binding fragment thereof, eg, secukinumab, weekly, every two weeks, every month [every four weeks], every other month, four times a year, twice a year or Includes annual administration. In some embodiments, the maintenance regimen uses monthly dosing (every 4 weeks). In some embodiments, the first dose of the maintenance regimen is delivered during weeks 4 or 16. In some embodiments, the maintenance regimen comprises administering a dose of about 150 mg to 300 mg, such as about 150 mg or about 300 mg, of an IL-17 antibody or antigen-binding fragment thereof, such as secukinumab.

  Delivery of an IL-17 antibody, such as secukinumab, during a loading, induction, and / or maintenance regimen is by subcutaneous route, eg, about 75 mg to about 300 mg (eg, about 50 mg, about 75 mg, about 100 mg, about 125 mg, about 150 mg). , About 175 mg, about 200 mg, about 225 mg, about 250 mg, about 275 mg, about 300 mg, about 325 mg), intravenous route, eg about 1 mg / kg to about 50 mg / kg (eg about 1 mg / kg, Delivery of a dosage of about 3 mg / kg, about 10 mg / kg, about 30 mg / kg, about 40 mg / kg, about 50 mg / kg, etc.), or any other route of administration (eg, intramuscular, im) Can be. In a preferred embodiment, the dose of IL-17 antibody is s. c. Delivered.

  In a preferred embodiment, the patient has about 150 mg to about 300 mg (e.g., about 150 mg or about 300 mg) with initial dosing at weeks 0, 1, 2 and 3 followed by monthly maintenance starting at 4 weeks. ) Dose of IL-17 antibody or antigen-binding fragment thereof, such as secukinumab, is administered by subcutaneous injection. In this regimen, dosing is performed during each week period, such as weeks 0, 1, 2, 3, 4, 8, 12, 16, and 20. The 300 mg dose may be given as two 150 mg subcutaneous injections.

  As used herein, a method of treating an autoimmune disease (eg, psoriasis, rheumatoid arthritis, ankylosing spondylitis, psoriatic arthritis) comprising 0, 1, 2, and 3 for patients in need thereof. A dose of about 150 mg to about 300 mg (e.g., about 150 mg or about 300 mg) of IL-17 antibody or antigen-binding fragment thereof, e.g., with initial administration at week and monthly maintenance administration starting at week 4 Comprising administering secukinumab by subcutaneous injection, wherein the IL-17 antibody or antigen-binding fragment thereof, eg secukinumab, is about 20 mg / ml to about 175 mg / ml (eg, about 25 mg / ml to about 150 mg / ml) of the present specification. An IL-17 antibody or antigen-binding fragment thereof (e.g., secukinumab), a buffer having a pH of about 5.2 to about 6.2, and Provided as part of a pharmaceutical composition comprising a methionine about 2.5～20MM, Liquid pharmaceutical compositions manner not restored from lyophilisate is disclosed.

  As used herein, an IL-17 antibody (eg, secukinumab) for the manufacture of a medicament for the treatment of an autoimmune disease (eg, psoriasis, rheumatoid arthritis, ankylosing spondylitis, psoriatic arthritis) in a patient. In use, the medicament comprises containers each having a headspace having an oxygen content of less than about 12% (eg, less than about 10%, less than about 8%, less than about 7%, less than about 6%, etc.), and Formulated to include a liquid pharmaceutical composition disposed in a container, wherein the composition is from about 20 mg / ml to about 175 mg / ml (eg, from about 25 mg / ml to about 150 mg / ml) herein. A liquid having a pH of about 5.2 to about 6.2, and about 2.5 to 20 mM methionine; Drug composition is not restored from the freeze-dried product, use is disclosed.

Kits comprising pharmaceutical products and liquid compositions The present disclosure also encompasses kits for treating various autoimmune diseases (eg, psoriasis). In a broad sense, such a kit includes at least one of the disclosed pharmaceutical product or liquid composition and instructions for use. The instructions disclose suitable techniques for providing the patient as part of a stable liquid composition dosing regimen. These kits are additional agents for treating autoimmune diseases (eg, psoriasis) for delivery (ie, simultaneous or sequential [before or after]) with the encapsulated liquid composition. The above) can also be contained.

  As used herein, a kit for the treatment of a patient having an autoimmune disease (eg, psoriasis), wherein a) the oxygen content is less than about 12% (eg, less than about 10%, less than about 8%, about A container having a head space that is less than 7%, such as less than about 6%), b) a liquid pharmaceutical composition disposed in said container, i) from about 20 mg / ml to about 175 mg / ml (eg, About 25 mg / ml to about 150 mg / ml) of an IL-17 antibody disclosed herein or an antigen-binding fragment thereof (eg, secukinumab); ii) a buffer having a pH of about 5.2 to about 6.2 And iii) a liquid pharmaceutical composition comprising about 2.5-20 mM methionine and not reconstituted from the lyophilizate; and c) a kit comprising instructions for administering the liquid pharmaceutical composition to a patient. The In some embodiments, the container is a pen, pre-filled syringe, auto-injector or vial.

General In some embodiments of the present disclosure, the IL-17 antibody or antigen-binding fragment thereof is selected from the group consisting of: a) Leu74, Tyr85, His86, Met87, Asn88, Val124, Thr125, Pro126, Ile127 IL-17 antibody that binds to an epitope of IL-17, including Val128, His129; b) an IL-17 antibody that binds to an epitope of IL-17, including Tyr43, Tyr44, Arg46, Ala79, Asp80; c) two matures Binds to an epitope of IL-17 homodimer with IL-17 protein chain, which epitope is on one chain Leu74, Tyr85, His86, Met87, Asn88, Val124, Thr125, Pro126, Ile127, Val128, H is129 binds to an epitope of an IL-17 homodimer with two mature IL-17 protein chains, comprising Tyr43, Tyr44, Arg46, Ala79, Asp80 on the other chain; d) IL-17 antibody containing Leu74, Tyr85, His86, Met87, Asn88, Val124, Thr125, Pro126, Ile127, Val128, His129 on one chain and Tyr43, Tyr44, Arg46, Ala79, Asp80 on the other chain there are about 100 to about 200 pM (e.g., Biacore (as measured by TM)) has a _{K D} of, IL-17 binding molecules having a in vivo half-life of about 23 to about 30 days; and e ) Selected from the group consisting of IL-17 antibodies including antibody: i) an immunoglobulin heavy chain variable domain comprising the amino acid sequence shown in SEQ ID NO: 8 _(V H); ii) an immunoglobulin light chain variable domain comprising the amino acid sequence shown in SEQ ID NO: 10 _{(V L} Iii) an immunoglobulin V _H domain comprising the amino acid sequence set forth in SEQ ID NO: 8 and an immunoglobulin _VL domain comprising the amino acid sequence set forth in SEQ ID NO: 10; iv) the supersets shown in SEQ ID NO: 1, SEQ ID NO: 2 and SEQ ID NO: 3 An immunoglobulin V _H domain comprising the variable regions in order; v) an immunoglobulin _VL domain comprising the hypervariable regions shown in SEQ ID NO: 4, SEQ ID NO: 5 and SEQ ID NO: 6 in order; vi) SEQ ID NO: 11, SEQ ID NO: 12 and An immunoglobulin V _H domain comprising in turn the hypervariable region shown in SEQ ID NO: 13; vii) SEQ ID NO: 1, SEQ ID NO: 2 And an immunoglobulin V _H domain comprising in turn the hypervariable region shown in SEQ ID NO: 3 and an immunoglobulin V _L domain comprising in turn the hypervariable region shown in SEQ ID NO: 4, SEQ ID NO: 5 and SEQ ID NO: 6; and viii) SEQ ID NO: 11, an immunoglobulin V _H domain comprising in turn the hypervariable regions shown in SEQ ID NO: 12 and SEQ ID NO: 13 and an immunoglobulin _VL domain comprising in turn the hypervariable regions shown in SEQ ID NO: 4, SEQ ID NO: 5 and SEQ ID NO: 6; ix) an immunoglobulin heavy chain comprising the amino acid sequence shown in SEQ ID NO: 15 (with or without C-terminal lysine); x) an immunoglobulin light chain comprising the amino acid sequence shown in SEQ ID NO: 14; xi) in SEQ ID NO: 15 An immunoglobulin heavy chain (with or without C-terminal lysine) containing the amino acid sequence shown and shown in SEQ ID NO: 14 An immunoglobulin light chain comprising the amino acid sequence. In some embodiments of the present disclosure, the IL-17 antibody or antigen-binding fragment thereof is a human antibody, preferably secukinumab.

  The details of one or more embodiments of the disclosure are set forth in the accompanying description above. Although any methods and materials similar or equivalent to those described herein can be used in the practice or testing of the present disclosure, the preferred methods and materials are now described. Other features, objects, and advantages of the disclosure will be apparent from the description and the claims. In this specification and the appended claims, the singular forms include the plural unless the context clearly dictates otherwise. Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this disclosure belongs. All patents and publications cited in this specification are incorporated by reference. The following examples are presented in order to more fully illustrate the preferred embodiments of the disclosure. These examples should in no way be construed as limiting the scope of the disclosed subject matter as defined by the appended claims.

  The examples and embodiments described herein are for illustrative purposes only, and various modifications and changes will be suggested to those skilled in the art in light of the spirit and purpose of this application and the accompanying It is understood that it is within the scope of the claims.

  These examples describe the development of a stable liquid composition of secukinumab. The data show that the composition pH and the choice of the second group of stabilizers had a major impact on the stability of the liquid composition. The data also shows the effect of headspace oxygen content, which affects the stability of the liquid composition. The effects on stability of antibody concentration, surfactant selection, first group stabilizer selection and buffer system selection were less. Thus, when considering variables that have a greater impact on stability, the disclosed pharmaceutical product includes a container (eg, PFS or vial) having a headspace in which the oxygen content is less than about 12%, as well as in the container. Wherein the composition has a pH of about 5.2 to about 6.2 and has a pH of about 20 mg / mL to 175 mg / mL, and about 2.5 to about Contains about 20 mM L-methionine. When considering variables that have large and small effects on stability, the disclosed pharmaceutical product includes a container (eg, PFS or vial) having a headspace in which the oxygen content is less than about 12%, as well as in said container Wherein the composition has a pH of about 5.2 to about 6.2, and sequukinumab; a buffer; a surfactant, a stabilizer, and about 2.5 to about 20 mM. Contains L-methionine.

  Based on the data disclosed below, preferred liquid compositions are about 25 mg / mL to about 165 mg / mL secukinumab, about 185 mM to about 225 mM trehalose, about 0.01% to about 0.03% polysorbate 80, about 2. 5 mM to about 20 mM L-methionine and about 10-30 mM histidine buffer (eg, about 20 mM histidine buffer) are included at a pH of about 5.8.

  A preferred liquid composition I comprises about 150 mg / mL secukinumab, about 200 mM trehalose, about 0.02% polysorbate 80, about 5 mM L-methionine and about 20 mM histidine buffer at a pH of about 5.8. A preferred pharmaceutical product I comprises said liquid composition I placed in a prefilled syringe (PFS).

  Another preferred liquid composition II comprises about 25 mg / mL secukinumab, about 225 mM trehalose, about 0.02% polysorbate 80, about 5 mM L-methionine and about 20 mM histidine buffer at a pH of about 5.8. A preferred pharmaceutical product II comprises said liquid composition II placed in a vial.

1.1 Part I-Detailed analysis of variables that have a greater impact on liquid state stability (headspace oxygen, pH and L-methionine)
1.1.1 Example 1: L-methionine The effect of several antioxidant stabilizers on secukinumab stability was characterized using a series of extensive analytical techniques.

  Initial studies evaluated various antioxidant stabilizers including tetrasodium EDTA sodium ascorbate, cysteine, sodium bisulfite and sodium citrate. None of these stabilized the molecule sufficiently, but there was a small stabilizing effect of tetrasodium EDTA and sodium citrate on the agglomerated products by SEC compared to compositions that did not contain antioxidant stabilizers. (Data not shown).

  In further studies, cysteine, tetrasodium EDTA and L-methionine stabilizers were evaluated at a concentration of 10 mM using the Doe approach with a secukinumab concentration of 150 mg / mL and compared to no stabilizer. The composition was loaded into PFS and placed in a 2-month stability study under conditions of long-term (5 ° C.), accelerated (25 ° C.) and stress (40 ° C.), and physical stability (AP-SEC, DLS, Of turbidity, visible and sub-visible particles by light darkening), chemical stability (purity by CEX, purity by RP-HPLC, color) and biological activity (activity by Cys-CEX, free SH groups) The indicators were evaluated. In addition, freeze thaw (-5 cycles from -20 ° C. to room temperature) and shaking stress (1 week at 150 rpm) were added to the composition filled in 2 mL vials.

  For secukinumab, L-methionine was found to be the best second group of stabilizers. This was demonstrated by purity by CEX and purity by RP-HPLC and higher purity levels as measured by lower turbidity levels and visible particle counts. Compared to the composition without stabilizer, significantly better stability was shown in the presence of L-methionine. Compositions with L-methionine were found to have lower levels of AP-SEC, more consistent DLS data, lower turbidity and pre-reduction by RP-HPLC after 8 weeks of stability at 25 ° C and 40 ° C accelerated conditions. It had less amount of main peak species. EDTA was disadvantageous due to the increase in pre-main peak species by AP-SEC, DLS, CEX and basic mutants by RP-HPLC. As demonstrated by various analytical methods, cysteine leads to an increase in almost all aggregation and degradation products.

  FIG. 1 lists selected quality attributes after storage under different conditions. Only L-methionine was observed to have a consistent stabilizing effect on secukinumab. Stabilizing effects were particularly observed with pre-main peak species by RP-HPLC (FIG. 1B) and AP-SEC (FIG. 1D). Further effects were also observed with turbidity and hydrodynamic radius due to DLS. The effect of different L-methionine concentrations on secukinumab quality attributes was evaluated in subsequent studies.

  FIG. 2 shows storage at 25 ° C. at 25 mg / mL secukinumab concentration and 225 mM trehalose concentration and 0.02% polysorbate 80 concentration in histidine buffer pH 5.8 in the presence and absence of L-methionine. The change of the pre-main peak species by RP-HPLC during is shown. The composition was filled into 2 mL vials and stored for up to 3 months under stress conditions. The black dashed line represents the linear fit to the value obtained with the composition containing 0 mM L-methionine, and the gray dashed line represents the linear fit to the value obtained with the composition containing 10 mM L-methionine. Represent. Clearly, reduced degradation kinetics were observed in the presence of L-methionine.

  A concentration-dependent effect was also observed for the composition containing 150 mg / mL secukinumab. Studies were performed with compositions containing trehalose at a concentration between 200 mM and 300 mM, polysorbate 80 between 0.01% and 0.04%, and 0 mM to 10 mM L-methionine. The composition was filled into 1 mL PFS and stored for up to 3 months under long-term, accelerated and stressed conditions. Physical (AP-SEC, DLS, subvisible and visible particles by light darkening, turbidity) and chemical (purity by CEX, purity by RP-HPLC, color) stability and biological activity of secukinumab Was monitored.

  FIG. 3 shows the pre-main peak species by RP-HPLC after storage for 6 months at 25 ° C. While the effects of trehalose and polysorbate 80 on degradation were negligible, a clearly reduced degradation level was observed in the presence of L-methionine. This effect was more pronounced when comparing secukinumab stability with and without L-methionine, but concentration dependence was also observed in the range of 2.5-10 mM L-methionine.

  After long-term storage (up to 30 months) with a composition containing 150 mg / mL secukinumab, 200 mM trehalose, 0.02% polysorbate 80 in histidine buffer pH 5.8, loaded in 1 mL PFS, L- The same stabilizing effect of methionine was observed. FIG. 4 shows pre-main peak species (B) by AP-SEC (A) and RP-HPLC during storage at 5 ° C. for up to 30 months. The black dashed line represents the linear fit to the value obtained with the composition containing 5 mM L-methionine, and the gray dashed line represents the linear fit to the value obtained with the composition containing 0 mM L-methionine. Represent. Clearly, reduced degradation kinetics were observed in the presence of L-methionine.

  A study evaluating the effect of L-methionine concentration (0-20 mM) on secukinumab stability (150 mg / mL, trehalose 200 mM, polysorbate 80 0.02%, histidine buffer pH 5.8) further confirms concentration dependence It was. Different compositions were loaded into PFS and stored for 13 months as well as 30 months (5 ° C only) under long term and accelerated conditions. Sectukinumab stability was assessed by a selected set of analytical techniques that were observed to show stability on previous screens (RP-HPLC purity, SEC purity, turbidity). From the turbidity measurements, no obvious trend could be concluded. However, the pre-main peak species by AP-SEC and RP-HPLC showed a clear dependence on L-methionine concentration. Although this effect was small under real-time storage conditions, a clear difference was observed at 25 ° C. (FIG. 5).

  After 13 months storage at 25 ° C., the level of SEC aggregates in the composition without L-methionine increased by 4.5% from a starting level of less than 1% at t0. With the addition of L-methionine in the composition, this increase in aggregate formation was reduced to 3.5% at 2.5 mM, 3.0% at 5 mM and 2.2% at 20 mM L-methionine. At 5 ° C., the difference between the composition without L-methionine and the composition with 20 mM L-methionine was only 0.3%. The pre-main peak species by RP-HPLC was a sample containing 0 mM L-methionine and increased from 9.1% to 42.7% during 13 months storage at 25 ° C. This increase in pre-main peak species by RP-HPLC was reduced to 39.4% at 2.5 mM, 37.8% at 5.0 mM and 34.5% in the sample containing 20 mM L-methionine. In summary, reduced levels of pre-main peak species by AP-SEC and RP-HPLC were observed during storage at 5 ° C. and 25 ° C. in PFS in the presence of L-methionine. A clearer difference was observed after storage at 25 ° C, but was detectable after storage at 5 ° C.

  Already at the level of 2.5 mM L-methionine, the degradation rate was clearly reduced compared to the composition without L-methionine. This was also confirmed in further studies comparing secukinumab stability in the presence of 0, 2.5 and 5.0 mM L-methionine. No difference in RP-HPLC purity, SEC purity and turbidity is observed between compositions containing 2.5 mM and 5.0 mM L-methionine after 24 months storage at the intended storage conditions. It was.

  Addition of L-methionine to the liquid antibody composition in the vial also reduced impurities due to AP-SEC and CE-SDS (non-reducing) (FIG. 6). Interestingly, a reduced L-methionine concentration dependence was observed with liquid antibody compositions in vials with 25 mg / mL secukinumab (FIG. 6), with antibody integrity and lower antibody concentration compositions and A lower concentration of L-methionine is suggested to be sufficient to maintain stability.

  Based on the combined data from the above experiments, a methionine concentration of at least 2.5 mM (preferably about 5 mM) is ideal for secukinumab liquid compositions and is superior to the other second group of stabilizers.

1.1.2 Example 2: Headspace oxygen content 1.1.2.1 Primary packaging-PFS:
Evaluate the effect of headspace oxygen content on secukinumab stability at a composition of 150 mg / mL secukinumab and with 200 mM trehalose, 5 mM L-methionine, 0.02% polysorbate 80 in histidine buffer pH 5.8 did. 1 mL PFS from various PFS suppliers was filled with the composition. Headspace oxygen content is measured to be between 13% and 15% (0.5 mL fill volume), or 3-4% (0.5 mL fill volume) / 7-7% (1.0 mL fill volume), respectively. It was done. Samples were stored for up to 6 months under prolonged, accelerated and stress conditions. Selected compositions were stored for up to 24 months under long term conditions. Sechukinumab stability is determined by SEC purity, RP-HPLC purity, CEX purity, CE-SDS (non-reducing) purity, turbidity, color, free SH groups, biological activity, sub-visible by photodarkening. And monitored by visible particles.

  The effect of headspace oxygen content on pre-main peak species and AP-SEC by RP-HPLC was observed under prolonged, accelerated and stress conditions. FIG. 7 shows AP-SEC during storage at 25 ° C. for up to 9 months. Apparently, PFS with a headspace oxygen content of 13-15% showed increased aggregation at 25 ° C. However, there was little absolute difference in aggregate levels compared to the headspace oxygen content under storage conditions of 2-8 ° C. (6 months data) (data not shown).

  The quality attributes of secukinumab (turbidity, purity by SEC, purity by RP-HPLC, purity by CE-SDS (non-reducing), free SH groups, biological activity, sub-visible particles by light darkening, visible particles, The effect of different headspace oxygen content levels on the color) ranging from 6% to 21% (i.e. no purge) is 12 months at 5 ° C and 6 months under accelerated conditions (25 ° C) and under stress conditions ( 40 ° C) for further evaluation during storage for 3 months. Studies were performed with a composition having 200 mM trehalose, 5 mM L-methionine, 0.02% polysorbate 80 in 150 mg / mL secukinumab and in histidine buffer pH 5.8. Samples were loaded into PFS and purged with a certified oxygen mixture to give the target headspace oxygen content.

  Over the storage time no change in turbidity is observed; no clear effect of headspace oxygen content on sub-visible particles, color and free SH groups due to light darkening is observed, and the difference between different headspace oxygen content samples Was within the range of method variation. The methionine concentration shows no relevant change during storage at 5 ° C. or 25 ° C. and is 4.9 mM (initial value 4.9-5.0 mM) after 12 months at 5 ° C., regardless of the headspace oxygen content. It was observed that there was.

  In contrast to our previous findings that showed a relatively large effect of headspace oxygen content on the agglomerated product by SEC, in this experiment, even the unpurged reference sample, even the intended storage conditions ( Only small changes were observed during storage for up to 12 months at 5 ° C. For different stability time points tested (storage up to 12 months at 2-8 ° C. and storage up to 6 months at 25 ° C.) between samples with different oxygen levels in the headspace, purity and aggregates by SEC No related differences were observed (Figure 8). In contrast, we noticed an increase in main purity by RP-HPLC with increasing headspace oxygen content. This was observed at 5 ° C. (after 12 months storage) (data not shown) and 25 ° C. (after 6 months storage) (FIG. 9). No new peak appeared.

1.1.2.2 Primary packaging-vials:
The composition was filled into 2 mL vials and stored for 12 months under refrigerated conditions and up to 3 months under accelerated and stressed conditions. Tables 5-7 show 25 mg / mL secukinumab concentration and 225 mM trehalose concentration and 0.02% polysorbate 80 concentration in histidine buffer pH 5.8 in the presence and absence of 5 mM L-methionine. Summarizes changes in pre-main peak species and SEC-AP by RP-HPLC during storage at 5 ° C, 25 ° C and 40 ° C.

  The effect of headspace oxygen content on the 25 mg / ml secukinumab vials was 5 months after 12 months (4.5% at 5% headspace oxygen content 7.1% at 20% headspace oxygen content, table 5) after 3 months at 25 ° C. (17.9% with 5% headspace oxygen content vs. 20.6% with 20% oxygen, see Table 6) and after 3 months at 40 ° C. (5 % Headspace oxygen content 40.6% vs. 20% headspace oxygen content 46.2%, see Table 6), seen by pre-main peak species by RP-HPLC. The same trend can be derived for AP-SEC after 3 months at 40 ° C. (1.8% with 5% headspace oxygen content versus 2.5% with 20% headspace oxygen content, see Table 6). Furthermore, L-methionine concentration has an additional effect when combined with a lower headspace oxygen content. For example, comparing pre-main peak species by RP-HPLC with a composition containing 5% oxygen headspace content data after 12 months of storage at 5 ° C., 4 of the composition with 5 mM L-methionine. Compared to 5% (Table 5), 6.1% (Table 7) was found in the composition containing no L-methionine. After 3 months at 25 ° C. (5-20% oxygen: 20.9-25.5% in the absence of L-methionine (Table 8) vs. 17.9-25. 5 in the presence of 5 mM L-methionine. 5% (Table 6)), and after 3 months at 40 ° C. (5% -20% oxygen: 44.6-50.9% (Table 8) vs. 5 mM L-methionine in the absence of L-methionine The same difference is seen with pre-main peak species by RP-HPLC, in the presence of 40.6-46.2% (Table 6)).

  Based on the above experiment, the headspace oxygen content is reduced to less than about 12% in enhancing the stability of the liquid composition in both PFS and vial (as assessed by pre-main peak species by RP-HPLC). Nitrogen purge is considered beneficial.

1.1.3 Example 3: Interaction between L-methionine concentration and headspace oxygen content Further studies evaluated the interaction between L-methionine concentration and headspace oxygen content. Compositions were prepared containing L-methionine in the range of 2.5-7.5 mM and a headspace oxygen content between 3-9%. The composition was loaded into PFS and stored for 6 months under prolonged and accelerated conditions. Relevant secukinumab quality attributes (purity by SEC, purity by RP-HPLC; purity by CEX, free SH groups, biological activity, sub-visible and visible particles by light darkening, turbidity and color of the solution), Monitoring was performed after 3 and 6 months of storage. FIG. 10 shows the purity by AP-SEC after 6 months storage at 25 ° C., corresponding to L-methionine and headspace oxygen content. When analyzed using purity by AP-SEC, no interaction was observed in the range tested.

  In another study, the effects of reduced headspace oxygen content and L-methionine concentration were evaluated at a secukinumab concentration of 150 mg / mL. The composition contained 270 mM mannitol, 0.04% polysorbate 80 and different L-methionine concentrations ranging from 0.15% to 2%. The composition was filled into 2 mL glass vials and either purged with nitrogen or not and stored for up to 6 months under prolonged, accelerated and stress conditions.

  FIG. 11 shows that after storage for up to 36 months in a composition containing 0.15% (10 mM), 1% (67 mM) or 2% (134 mM) L-methionine and nitrogen or air headspace. Represents pre-main peak species by RP-HPLC. As previously observed, pre-main peak species by RP-HPLC were at lower levels in compositions containing higher amounts of L-methionine. When the headspace was purged with nitrogen, the same composition showed a lower level of pre-main peak species by RP-HPLC.

  Based on combined data from various experiments using vials and PFS as primary packaging, a headspace oxygen content of less than about 12% in combination with an L-methionine concentration of at least about 2.5 mM was found in the liquid composition of secukinumab. Ideal.

1.1.4 Example 4: pH
The effect of pH on secukinumab stability was initially evaluated at a concentration of 10 mg / ml in 100 mM citrate / sodium phosphate buffer containing 90 mM sodium chloride in the pH range between 4.0 and 7.5. did. Samples were stored at 5 ° C. and 40 ° C. for 3 weeks. In parallel, secukinumab stability after 5 freeze-thaws from ≦ −60 ° C. to room temperature was monitored.

  The optimum pH for secukinuma depended on the degradation pathway analyzed. Aggregation and proteolysis were determined by SEC purity, SDS-PAGE (reduction) purity, and the average molecular weight by LLS was minimal at pH 5.7-6.2, but the optimum pH for CEX purity was pH 5. 3. Active secukinumab contains one free cysteine residue on each light chain, and thus 2 moles of thiol group are expected per mole of secukinumab. Since reduced levels of free SH groups correlate with the loss of biological activity of secukinumab, free SH groups were quantified using the Elman reagent based method. A slightly lower value of 1.94 moles per mole was observed at pH 4.3 only. The freeze-thaw resistance of secukinumab was monitored by SEC purity, and the average molecular weight by LLS was highest at pH 5.3 to 5.7. A pH of 5.8 was chosen to further formulate secukinumab.

  Further studies on the effect of pH on secukinumab stability were performed with PFS using the Doe approach. The effect of pH within the range of 5.2 to 5.8 was evaluated at a secukinumab concentration of 150 mg / mL. The composition was placed in a 2-month stability study under conditions of long-term (5 ° C.), accelerated (25 ° C.) and stress (40 ° C.), physical stability (AP-SEC, DLS, turbidity, light darkness Visual and sub-visible particles), chemical stability (purity by CEX, purity by RP-HPLC, color) and biological activity (activity by Cys-CEX, free SH groups). In addition, freeze thaw (-5 cycles from -20 ° C. to room temperature) and shaking stress (1 week at 150 rpm) were added to the composition filled in 2 mL vials. A pH value within the investigated range was found to significantly affect secukinumab stability (AP-SEC, DP-SEC, DLS, basic variants by CEX, purity by RP-HPLC). Results from previous studies confirmed that pH 5.8 is ideal (pre-main peak species by AP-SEC, DP-SEC and RP-HPLC) (Figure 12).

  Further evaluation of pH effects with compositions containing secukinumab at a concentration of 150 mg / mL, trehalose 200 mM, L-methionine in the range 2.5-7.5 mM and headspace oxygen content between 3 and 9% did. The pH of the histidine buffer varied between 5.4 and 6.2. The composition was loaded into PFS and stored for 6 months under prolonged and accelerated conditions. Relevant secukinumab quality attributes (purity by SEC, purity by RP-HPLC; purity by CEX, free SH groups, biological activity, sub-visible particles by light darkening, visible particles, solution turbidity and color) Monitoring was performed after 3 and 6 months of storage. FIG. 13 represents the effect of pH on secukinumab quality attributes after storage at 5 ° C. Increased turbidity, acidic variants with AP-SEC and CEX and reduced purity with SEC were observed at higher pH values, further confirming the observations from the initial screening.

  Based on combined data from various experiments, a pH range of about 5.2 to about 6.2 is ideal for liquid compositions of secukinumab.

1.2 Part 2-Detailed analysis of excipients with less impact on liquid state stability (stabilizers, surfactants and buffers)
1.2.1 Example 5: Stabilizer selection has little effect on stability Long term storage conditions and soluble and insoluble aggregate formation of secukinumab during storage under accelerated and stress conditions (AP-SEC, SDS-PAGE) Purity, light scattering technology), chemical stability (RP-HPLC purity, CEX purity, color) and biological activity (Cys-CEX activity, free SH groups, biological activity) different stability Development of initial compositions for liquid dosage forms with emphasis on drug evaluation.

  Stabilizers were divided into three different classes: The first group included nonionic (mannitol, trehalose dihydrate) and ionic (sodium chloride and arginine hydrochloride) stabilizers. All stabilizers of the first group provided advantages over those without stabilizers. However, nonionic stabilizers (trehalose and mannitol) were observed to better stabilize the molecule as observed by lower aggregate levels and higher activity with Cys-CEX.

  Based on the conclusions from the initial composition development studies, further studies were performed at PFS using the DoE approach. The effect of the first stabilizer group (glycine, mannitol, trehalose dihydrate, sodium chloride) was evaluated. The composition is filled into pre-filled syringes and placed in a 2-month stability study under conditions of long-term, accelerated and stressed, physical stability (AP-SEC, DLS, turbidity, visible and subtracted by light darkening) Visible particles), chemical stability (CEX purity, RP-HPLC purity, color) and biological activity (Cys-CEX activity, free SH groups) indicators were evaluated. In addition, freeze thaw (-5 cycles from -20 ° C. to room temperature) and shaking stress (1 week at 150 rpm) were added to the composition filled in 2 mL vials. With respect to stabilizer class I, observations from previous screens were confirmed: 1) all stabilizers of the first group provided advantages over those without stabilizers; and 2) nonionic stabilizers were It was found to be a better stabilizer for secukinumab protein (FIG. 14). This was particularly noticeable with SEC purity, RP-HPLC purity and DLS polydispersity. No related effects were observed when comparing different nonionic stabilizers.

  Next, we identified the ideal concentration of stabilizer class I (trehalose dihydrate, 200-300 mM). Samples were loaded into PFS and stored for up to 3 months under prolonged, accelerated and stress conditions. Physical (AP-SEC, DLS, sub-visible particles by light darkening, visible particles, turbidity) and chemical (purity by CEX, purity by RP-HPLC, color) stability and biological activity of secukinumab Was monitored. At various trehalose concentrations, no related differences in secukinumab quality attributes were observed (Figure 3).

1.2.2 Example 6: Selection of surfactant has little effect on stability Long-term storage conditions and soluble and insoluble aggregate formation of secukinumab during storage under accelerated and stress conditions (AP-SEC, SDS- PAGE purity, light scattering technology), chemical stability (RP-HPLC purity, CEX purity, color) and biological activity (Cys-CEX activity, free SH groups, biological activity) different Development of an initial composition for a 150 mg / ml liquid composition with an emphasis on the evaluation of excipients (eg, stabilizers and surfactants). Excipients were divided into three different classes: Group III contained surfactant polysorbates 20 and 80. During static storage, no difference was observed between polysorbates 20 and 80 at a concentration of 0.04% compared to no surfactant.

  Based on the conclusions from the initial composition development studies, further studies were performed at PFS using the DoE approach. The effect of surfactant (polysorbate 20, polysorbate 80, poloxamer 188, none) was evaluated. The composition is loaded into PFS and placed in a 2-month stability study under long-term, accelerated and stressed conditions, physical stability (AP-SEC, DLS, turbidity, visible and sub-visible particles by light darkening ), Chemical stability (CEX purity, RP-HPLC purity, color) and biological activity (Cys-CEX activity, free SH groups) indicators. In addition, freeze thaw (-5 cycles from -20 ° C. to room temperature) and shaking stress (1 week at 150 rpm) were added to the composition filled in 2 mL vials. The presence of surfactant was beneficial as observed by the lower turbidity levels and the number of visible and sub-visible particles by light darkening. However, there was only a weak effect of the surfactant type (FIG. 15).

  We next identified the ideal concentration of surfactant group III (polysorbate 80 0.01-0.04 (w / v)%). Samples were loaded into PFS and stored for up to 3 months under prolonged, accelerated and stress conditions. Physical (AP-SEC, DLS, sub-visible particles by light darkening, visible particles, turbidity) and chemical (purity by CEX, purity by RP-HPLC, color) stability and biological activity of secukinumab Was monitored. No apparent effect of polysorbate 80 concentration on secukinumab quality attributes was observed during static storage (Figure 3) and after 1 week of shaking at 150 rpm. At higher surfactant concentrations, there was a slight increase in sub-visible particles due to polydispersity by DLS and light darkening. Therefore, in order to maintain a safety margin for the lowest evaluation concentration, the concentration of polysorbate 80 was defined as 0.02%.

1.2.3 Example 5: Selection of buffer has little impact on stability Using the DOE approach, the effect of buffer type (citric acid, histidine, succinic acid, acetic acid) was evaluated with PFS . The composition is loaded into PFS and placed in a 2-month stability study under long-term, accelerated and stressed conditions, physical stability (AP-SEC, DLS, turbidity, visible and sub-visible particles by light darkening ), Chemical stability (CEX purity, RP-HPLC purity, color) and biological activity (Cys-CEX activity, free SH groups) indicators. In addition, freeze thaw (-5 cycles from -20 ° C. to room temperature) and shaking stress (1 week at 150 rpm) were added to the composition filled in 2 mL vials. No related effects of buffer type were observed. FIG. 16 shows the selected quality attributes.

1.2.4 Example 6: Antibody concentration within the tested range has little effect on stability The effect of secukinumab concentration on liquid composition quality attributes was evaluated in the range of 124.5 to 175.5 mg / mL . The composition also contained 200 mM trehalose, 5 mM L-methionine and 0.02% polysorbate 80 in histidine buffer pH 5.8. The composition was loaded into PFS and stored for 6 months under prolonged and accelerated conditions. Relevant secukinumab quality attributes (purity by SEC, purity by RP-HPLC; purity by CEX, free SH groups, biological activity, sub-visible particles by light darkening, visible particles, solution turbidity and color) Monitoring was performed after 3 and 6 months of storage. Within the range of 25 mg / ml to 150 mg / ml, no associated effect of secukinumab concentration on liquid composition quality attributes was observed (data not shown).

1.3 Part 3-Properties of the preferred final commercial composition The preferred pharmaceutical product of secukinumab is 150 mg / ml secukinumab, 200 mM trehalose, 0.02% polysorbate 80 and 5 mM L-methionine in 20 mM histidine buffer pH 5.8. Liquid composition, which is provided in PFS. At the initial fill and end, the headspace in the PFS has an oxygen content of less than 12%. These pharmaceutical products have excellent shelf life and overall stability.

  Various batches of secukinumab pharmaceutical in PFS (150 mg / ml secukinumab, 200 mM trehalose dihydrate, 20 mM L-histidine / L-histidine hydrochloride monohydrate, 5 mM L-methionine, 0.02% polysorbate 80 (w / v%), pH 5.8) stability test was carried out. Tests for storage of up to 24 months under long-term storage conditions (2-8 ° C), storage of up to 6 months under accelerated storage conditions (25 ° C), and storage of up to 6 months under temperature stress conditions (30 ° C) The results are shown in Tables 9-11 below. Protects against light based on provided stability data, up to 24 months of real-time data for secukinumab 150 mg / 1 ml solution with prefilled syringe (PFS) and up to 36 months of stability data generated during development (bulk syringe) However, a shelf life of 24 months is proposed for secukinumab 150 mg / 1 ml solution in PFS commercial products when freezing is prevented and stored at long term conditions of 5 ° C. ± 3 ° C.

The present application relates to the following inventions.
(1) a. A container having a headspace in which the oxygen content is less than about 12%, and
b. A liquid pharmaceutical composition having a pH of about 5.2 to about 6.2, disposed in said container,
i. About 20 mg / ml to about 175 mg / ml secukinumab; and
ii. About 2.5 to about 20 mM L-methionine
A liquid pharmaceutical composition which is not reconstituted from a lyophilizate
Containing pharmaceutical products.
(2) The pharmaceutical product according to (1) above, wherein the concentration of methionine is about 2.5 mM, about 5 mM, about 10 mM, or about 20 mM.
(3) The pharmaceutical product according to (2) above, wherein the concentration of methionine is about 5 mM.
(4) The pharmaceutical product according to any one of (1) to (3), wherein the oxygen content in the head space is less than about 10%.
(5) The pharmaceutical product according to any one of (1) to (4), wherein the oxygen content in the head space is less than about 8%.
(6) The pharmaceutical product according to any one of (1) to (5) above, wherein the oxygen content in the head space is less than about 6%.
(7) The pharmaceutical product according to any one of (1) to (6) above, wherein the liquid pharmaceutical composition has a pH of about 5.8.
(8) The pharmaceutical product according to any one of (1) to (7) above, wherein the concentration of secukinumab is about 25 mg / ml or about 150 mg / ml.
(9) Any of (1) to (8) above, wherein the liquid pharmaceutical composition further comprises a buffer selected from the group consisting of a histidine buffer, a citrate buffer, an acetate buffer and a succinate buffer A pharmaceutical product according to claim 1.
(10) The pharmaceutical product according to (9) above, wherein the buffer is at a concentration of about 10 mM to about 30 mM.
(11) The pharmaceutical product according to (10) above, wherein the buffer is a histidine buffer having a concentration of about 20 mM.
(12) The pharmaceutical product according to any one of (1) to (11) above, wherein the liquid pharmaceutical composition further comprises a surfactant selected from polysorbate and poloxamer.
(13) The pharmaceutical product according to any one of (1) to (12), wherein the surfactant is polysorbate 80, polysorbate 20, or poloxamer 188.
(14) The pharmaceutical product according to (13) above, wherein the surfactant is polysorbate 80 at a concentration of about 0.01% (w / v) to about 0.04% (w / v).
(15) The pharmaceutical product according to (14) above, wherein the concentration of polysorbate 80 is about 0.02% (w / v).
(16) The pharmaceutical product according to (13) above, wherein the surfactant is polysorbate 20 at a concentration of about 0.02% (w / v).
(17) The liquid pharmaceutical composition according to any one of (1) to (16), further comprising a stabilizer selected from the group consisting of mannitol, sodium chloride, trehalose, arginine HCl, and glycine. Pharmaceutical products.
(18) The pharmaceutical product according to (17), wherein the stabilizer is trehalose at a concentration of about 180 mM to about 300 mM.
(19) The pharmaceutical product according to (18) above, wherein the concentration of trehalose is about 200 mM or about 225 mM.
(20) The pharmaceutical product according to any one of (1) to (19), wherein the container is a cartridge, a syringe, a pen, or a vial.
(21) a. A container having a headspace in which the oxygen content is less than about 6%, and
b. A liquid pharmaceutical composition disposed in the container, comprising about 25 mg / mL to about 150 mg / mL secukinumab, about 10 mM to about 30 mM histidine pH 5.8, about 200 mM to about 225 mM trehalose, about 0.02% A liquid pharmaceutical composition comprising polysorbate 80 and from about 2.5 mM to about 20 mM methionine and not reconstituted from a lyophilizate
Containing pharmaceutical products.
(22) The pharmaceutical product according to (21) above, comprising about 25 mg / ml secukinumab and about 225 mM trehalose.
(23) The pharmaceutical product according to (21) above, comprising about 150 mg / ml secukinumab and about 200 mM trehalose.
(24) The liquid composition is
a. At least about 86% purity by RP-HPLC after 6 months storage at 2-8 ° C., at least about 76% purity by RP-HPLC after 6 months storage at 25 ° C./60% RH, and / or 30 ° C. / At least about 60% purity by RP-HPLC after 6 months storage at 75% RH;
b. At least about 77% purity by CEX after 6 months storage at 2-8 ° C., at least about 62% purity by CEX after 6 months storage at 25 ° C./60% RH, and / or at 30 ° C./75% RH A purity of at least about 50% by CEX after 6 months of storage;
c. At least about 98% purity by SEC after 6 months storage at 2-8 ° C., at least about 96% purity by SEC after 6 months storage at 25 ° C./60% RH, and / or at 30 ° C./75% RH A purity of at least about 94% by SEC after 6 months of storage;
d. At least about 97% purity by CE-SDS (non-reducing conditions) after 6 months storage at 2-8 ° C., at least about 95 by CE-SDS (non-reducing conditions) after 6 months storage at 25 ° C./60% RH Purity and / or purity of at least about 94% (preferably at least about 92%) by CE-SDS (non-reducing conditions) after 6 months storage at 30 ° C./75% RH;
e. Less than about 0.57% impurities by CE-SDS (reducing conditions) after 6 months storage at 2-8 ° C., about 1. by CE-SDS (reducing conditions) after 6 months storage at 25 ° C./60% RH. Less than 1% impurities and / or less than about 1.9% impurities by CE-SDS (reducing conditions) after 6 months storage at 30 ° C./75% RH; and / or
f. Relative biological activity of at least about 88% by inhibition of IL-6 release from C-20 / A4 chondrocytes after 24 months storage at 2-8 ° C., after 6 months storage at 25 ° C./60% RH At least about 94% relative biological activity by inhibiting IL-6 release from chondrocytes, and / or at least by inhibiting IL-6 release from chondrocytes after 6 months storage at 30 ° C./75% RH About 85% relative biological activity
The pharmaceutical product according to any one of (1) to (23), wherein
(25) The liquid composition is
a. A purity of at least about 84% by RP-HPLC after storage at 2-8 ° C. for 24 months;
b. A purity of at least about 73% by CEX after 24 months storage at 2-8 ° C;
c. A purity of at least about 97% by SEC after 24 months storage at 2-8 ° C;
d. At least about 97% purity by CE-SDS (non-reducing conditions) after 24 months storage at 2-8 ° C; and / or
e. Less than about 0.91% impurities due to CE-SDS (non-reducing conditions) after 24 months storage at 2-8 ° C
The pharmaceutical product according to any one of (1) to (24), wherein
(26) A method for reducing the oxidation of secukinumab,
a. Having a pH of about 5.2 to about 6.2;
i. About 25 mg / ml to about 150 mg / ml secukinumab; and
ii. About 2.5 mM to about 20 mM L-methionine
Preparing a liquid composition comprising
b. Placing the liquid composition in a container having a headspace; and
c. Adjusting the oxygen content in the headspace to about 12% or less;
Including methods.
(27) The method according to (26), wherein the adjusting step c) is performed by purging the head space using an inert gas.
(28) The method according to (27) above, wherein the inert gas is nitrogen or argon.
(29) The method according to (26) above, wherein the concentration of methionine is about 2.5 mM, about 5 mM, about 10 mM, or about 20 mM.
(30) The method according to (29) above, wherein the concentration of methionine is about 5 mM.
(31) The method according to (26), wherein the oxygen content in the head space is adjusted to less than about 10%.
(32) The method of (26) above, wherein the oxygen content in the headspace is adjusted to less than about 8%.
(33) The method of (26) above, wherein the oxygen content in the headspace is adjusted to less than about 6%.
(34) The method according to (26) above, wherein the liquid composition has a pH of about 5.8.
(35) The method according to (26) above, wherein the concentration of secukinumab is about 25 mg / ml or about 150 mg / ml.
(36) about 25 mg / mL to about 150 mg / mL secukinumab, about 10 mM to about 30 mM histidine pH 5.8, about 200 mM to about 225 mM trehalose, about 0.02% polysorbate 80 and about 2.5 mM to about 20 mM methionine A liquid pharmaceutical composition that has not been reconstituted from a lyophilizate.

Claims (36)

a. A container having a headspace in which the oxygen content is less than about 12%, and
b. A liquid pharmaceutical composition having a pH of about 5.2 to about 6.2, disposed in said container,
i. About 20 mg / ml to about 175 mg / ml secukinumab; and ii. A pharmaceutical product comprising a liquid pharmaceutical composition comprising about 2.5 to about 20 mM L-methionine and not reconstituted from a lyophilizate.   2. The pharmaceutical product of claim 1, wherein the concentration of methionine is about 2.5 mM, about 5 mM, about 10 mM or about 20 mM.   The pharmaceutical product according to claim 2, wherein the concentration of methionine is about 5 mM.   The pharmaceutical product according to any one of the preceding claims, wherein the oxygen content in the headspace is less than about 10%.   The pharmaceutical product according to any one of the preceding claims, wherein the oxygen content in the headspace is less than about 8%.   The pharmaceutical product according to any one of the preceding claims, wherein the oxygen content in the headspace is less than about 6%.   The pharmaceutical product according to any one of the preceding claims, wherein the liquid pharmaceutical composition has a pH of about 5.8.   The pharmaceutical product according to any one of the preceding claims, wherein the concentration of secukinumab is about 25 mg / ml or about 150 mg / ml.   The pharmaceutical product according to any one of the preceding claims, wherein the liquid pharmaceutical composition further comprises a buffer selected from the group consisting of histidine buffer, citrate buffer, acetate buffer and succinate buffer. .   10. The pharmaceutical product of claim 9, wherein the buffer is at a concentration of about 10 mM to about 30 mM.   11. A pharmaceutical product according to claim 10, wherein the buffer is a histidine buffer at a concentration of about 20 mM.   The pharmaceutical product according to any one of the preceding claims, wherein the liquid pharmaceutical composition further comprises a surfactant selected from polysorbates and poloxamers.   The pharmaceutical product according to any one of the preceding claims, wherein the surfactant is polysorbate 80, polysorbate 20 or poloxamer 188.   14. The pharmaceutical product of claim 13, wherein the surfactant is polysorbate 80 at a concentration of about 0.01% (w / v) to about 0.04% (w / v).   15. A pharmaceutical product according to claim 14, wherein the concentration of polysorbate 80 is about 0.02% (w / v).   14. The pharmaceutical product of claim 13, wherein the surfactant is polysorbate 20 at a concentration of about 0.02% (w / v).   The pharmaceutical product according to any one of the preceding claims, wherein the liquid pharmaceutical composition further comprises a stabilizer selected from the group consisting of mannitol, sodium chloride, trehalose, arginine HCl, and glycine.   18. A pharmaceutical product according to claim 17, wherein the stabilizer is trehalose at a concentration of about 180 mM to about 300 mM.   19. The pharmaceutical product according to claim 18, wherein the concentration of trehalose is about 200 mM or about 225 mM.   The pharmaceutical product according to any one of the preceding claims, wherein the container is a cartridge, syringe, pen or vial. a. A container having a headspace in which the oxygen content is less than about 6%, and
b. A liquid pharmaceutical composition disposed in the container, comprising about 25 mg / mL to about 150 mg / mL secukinumab, about 10 mM to about 30 mM histidine pH 5.8, about 200 mM to about 225 mM trehalose, about 0.02% A pharmaceutical product comprising a liquid pharmaceutical composition comprising polysorbate 80 and from about 2.5 mM to about 20 mM methionine and not reconstituted from a lyophilizate.   24. The pharmaceutical product of claim 21, comprising about 25 mg / ml secukinumab and about 225 mM trehalose.   24. The pharmaceutical product of claim 21, comprising about 150 mg / ml secukinumab and about 200 mM trehalose. The liquid composition is
a. At least about 86% purity by RP-HPLC after 6 months storage at 2-8 ° C., at least about 76% purity by RP-HPLC after 6 months storage at 25 ° C./60% RH, and / or 30 ° C. / At least about 60% purity by RP-HPLC after 6 months storage at 75% RH;
b. At least about 77% purity by CEX after 6 months storage at 2-8 ° C., at least about 62% purity by CEX after 6 months storage at 25 ° C./60% RH, and / or at 30 ° C./75% RH A purity of at least about 50% by CEX after 6 months of storage;
c. At least about 98% purity by SEC after 6 months storage at 2-8 ° C., at least about 96% purity by SEC after 6 months storage at 25 ° C./60% RH, and / or at 30 ° C./75% RH A purity of at least about 94% by SEC after 6 months of storage;
d. At least about 97% purity by CE-SDS (non-reducing conditions) after 6 months storage at 2-8 ° C., at least about 95 by CE-SDS (non-reducing conditions) after 6 months storage at 25 ° C./60% RH Purity and / or purity of at least about 94% (preferably at least about 92%) by CE-SDS (non-reducing conditions) after 6 months storage at 30 ° C./75% RH;
e. Less than about 0.57% impurities by CE-SDS (reducing conditions) after 6 months storage at 2-8 ° C., about 1. by CE-SDS (reducing conditions) after 6 months storage at 25 ° C./60% RH. Less than 1% impurities and / or less than about 1.9% impurities by CE-SDS (reducing conditions) after 6 months storage at 30 ° C./75% RH; and / or f. Relative biological activity of at least about 88% by inhibition of IL-6 release from C-20 / A4 chondrocytes after 24 months storage at 2-8 ° C., after 6 months storage at 25 ° C./60% RH At least about 94% relative biological activity by inhibiting IL-6 release from chondrocytes, and / or at least by inhibiting IL-6 release from chondrocytes after 6 months storage at 30 ° C./75% RH The pharmaceutical product according to any one of the preceding claims, which maintains a relative biological activity of about 85%. The liquid composition is
a. A purity of at least about 84% by RP-HPLC after storage at 2-8 ° C. for 24 months;
b. A purity of at least about 73% by CEX after 24 months storage at 2-8 ° C;
c. A purity of at least about 97% by SEC after 24 months storage at 2-8 ° C;
d. At least about 97% purity by CE-SDS (non-reducing conditions) after 24 months storage at 2-8 ° C .; and / or e. 8. The pharmaceutical product according to any one of the preceding claims, which retains less than about 0.91% impurities by CE-SDS (non-reducing conditions) after storage at 2-8 [deg.] C for 24 months. A method for reducing the oxidation of secukinumab,
a. Having a pH of about 5.2 to about 6.2;
i. About 25 mg / ml to about 150 mg / ml secukinumab; and ii. Preparing a liquid composition comprising about 2.5 mM to about 20 mM L-methionine;
b. Placing the liquid composition in a container having a headspace; and c. Adjusting the oxygen content in the headspace to about 12% or less.   27. The method of claim 26, wherein conditioning step c) is performed by purging the headspace using an inert gas.   28. The method of claim 27, wherein the inert gas is nitrogen or argon.   27. The method of claim 26, wherein the concentration of methionine is about 2.5 mM, about 5 mM, about 10 mM, or about 20 mM.   30. The method of claim 29, wherein the concentration of methionine is about 5 mM.   27. The method of claim 26, wherein the oxygen content in the headspace is adjusted to less than about 10%.   27. The method of claim 26, wherein the oxygen content in the headspace is adjusted to less than about 8%.   27. The method of claim 26, wherein the oxygen content in the headspace is adjusted to less than about 6%.   27. The method of claim 26, wherein the liquid composition has a pH of about 5.8.   27. The method of claim 26, wherein the concentration of secukinumab is about 25 mg / ml or about 150 mg / ml.   Contains about 25 mg / mL to about 150 mg / mL secukinumab, about 10 mM to about 30 mM histidine pH 5.8, about 200 mM to about 225 mM trehalose, about 0.02% polysorbate 80 and about 2.5 mM to about 20 mM methionine, lyophilized A liquid pharmaceutical composition that has not been reconstituted.